Wash monitor and method of use

ABSTRACT

The present disclosure provides a monitoring device comprising a test composition, a test element comprising a test portion to which the test composition is releasably adhered, a detection reagent, and a container comprising a first end with an opening and a second end opposite the first end. The test composition comprises a predetermined quantity of tracer analyte. The container is configured to receive the test portion and configured to be operationally coupled to an analytical instrument. The tracer analyte and the detection reagent each are capable of participating in one or more chemical reaction that results in the formation of a detectable product. A method of using the monitoring device to assess the efficacy of a washing process is also provided.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional PatentApplication Nos. 61/710,829, filed on Oct. 8, 2012 and 61/773,854, filedMar. 7, 2013, which are incorporated herein by reference in theirentirety.

BACKGROUND

Hospitals and clinics frequently rely on washing equipment and processesto remove biological soil from reusable medical instruments and devices.In addition, the solvent used in the washing processes may containchemical and/or enzymes to facilitate the removal and/or disinfection ofthe biological soil. In operation, the washing equipment can fail toadequately clean the instruments and devices due to one or more of avariety of reasons including, for example, washing the objects at anunacceptably low temperature and providing an inadequate volume and/orvelocity of solvent to the washing process. In addition, the washsolvent can fail to adequately clean the instruments and devices due toone or more of a variety of reasons including, for example, loss ofchemical and/or enzyme activity due to aging and improper dilution ofactive ingredients (e.g., chemicals or enzymes) in the wash solvent.

Disposable wash monitors are used for monitoring the efficacy of washprocesses in washer-disinfector equipment, for example. A wash monitortypically includes a test soil disposed on a surface of an object thatis placed into a washing machine. The test soil may comprise biologicalmolecules such as, for example, human or animal red blood cells,protein, and fat. The monitor also includes a detectable marker (e.g., apigment or dye) that can be observed to determine whether the washingmachine meets minimum requirements for impinging a wash solution againstan object and/or to determine whether the wash solution meets minimumrequirements for chemical and/or enzymatic treatment of the object to becleaned.

Although a variety of wash monitors are available to assess the efficacyof a washing process, there remains a need for improved wash monitors.

SUMMARY

The present disclosure generally relates to an article to assess theefficacy of a washing process and a method of use thereof. The inventivearticle is a modification of an existing article that is typically usedto detect a chemical analyte in a sample. The existing article includesa sample acquisition device (e.g., a swab) to obtain a sample with anunknown quantity of the chemical analyte. The inventive articledeliberately adulterates the sample acquisition device such that itcomprises a predetermined amount of the chemical analyte releasablyadhered thereto, thus destroying its utility for its originally-intendedpurpose. In the inventive method, the operator exposes the adulteratedsample acquisition device (hereinafter, “test element”) to a washingprocess and subsequently measures the quantity of chemical analyte(hereinafter, “tracer analyte”), if any, remaining on the test element.In addition, the present disclosure provides a test element with acomposition comprising the tracer analyte releasably adhered thereto anda system for assessing the efficacy of a washing process.

In one aspect, the present disclosure provides a monitoring device. Themonitoring device can comprise a test composition comprising apredetermined quantity of tracer analyte, a test element comprising atest portion to which the test composition is releasably adhered, adetection reagent, and a container comprising a first end with anopening and a second end opposite the first end. The container isconfigured to receive the test portion and configured to beoperationally coupled to an analytical instrument. The tracer analyteand the detection reagent each are capable of participating in one ormore chemical reaction that results in the formation of a detectableproduct. In any embodiment, the test portion can comprise at least onerecessed area, wherein the test composition is adhered in the recessedarea. In any of the above embodiments, the test composition further cancomprise a polymeric binder. In any of the above embodiments, the traceranalyte can be selected from the group consisting of a plurality ofviable microorganisms or a biomolecule associated therewith, an acid, abase, a nucleotide, a protein, a nucleic acid, a carbohydrate, orhemoglobin. In any of the above embodiments, the test portion cancomprise a porous fibrous nonwoven matrix, wherein the plurality ofviable microorganisms is disposed on or in the fibrous nonwoven matrix.

In any of the above embodiments, the monitoring device further cancomprise a frangible seal, wherein a receiving chamber is disposed on afirst side of the frangible seal proximate the opening and a cuvettechamber is disposed on a second side of the frangible seal distal theopening. In any of the above embodiments, container can include a firstsolvent disposed therein. In any of the above embodiments, the testelement further can comprise a reservoir with a second solvent disposedtherein, a hollow stem, and a liquid flow regulator capable of placingthe reservoir in fluid communication with the hollow stem. In any of theabove embodiments, the monitoring device further can comprise a securalelement.

In another aspect, the present disclosure provides a method of assessingthe efficacy of a washing process. The method can comprise exposing thetest portion of a monitoring device according to any one of the aboveembodiments to the washing process; after exposing the test portion tothe washing process, contacting the test portion with the detectionreagent in the container; and using the analytical instrument to detecta presence or an absence of the detectable product, wherein the presenceof the detectable product indicates a presence of tracer analyte on thetest portion after exposing the test portion to the washing process. Inany embodiment of the method, exposing the test portion to the washingprocess can comprise placing the test portion into an automated washerand performing at least a portion of an automated wash cycle while thetest portion is disposed in the automated washer. In any of the aboveembodiments of the method, using the analytical instrument to detect apresence or an absence of the detectable product can comprise using theanalytical instrument to measure a quantity of the tracer analyte. Inany of the above embodiments of the method, exposing the test portion ofa monitoring device can comprise exposing the test portion of aplurality of monitoring devices, wherein the method further can comprisepositioning a first monitoring device at a first predefined location inthe automated washer and positioning a second monitoring device at asecond predefined location in the automated washer. In any of the aboveembodiments, the method further can comprise comparing at least onemeasured quantity of the tracer analyte to a predefined standard.

In any of the above embodiments, the method further can compriseassociating a first datum related to a quantity of tracer analytedetected in a sample with a second datum related to other informationrelated to the sample and electronically storing the associated firstand second data. In some embodiments, the second datum can compriseinformation selected from the group consisting of a date, a time, awashing apparatus, an operator, an instrument to be washed, and acombination of two or more of any of the foregoing test data. In any ofthe above embodiments, the method further can comprise the step ofplacing the test portion of the monitoring device in a receiverconfigured to restrict fluidic accessibility to the test portion. In anyof the above embodiments of the method, placing the test portion of themonitoring device in a receiver configured to restrict fluidicaccessibility to the test portion can comprise placing the test portioninto an interior space of a lumened object.

In yet another aspect, the present disclosure provides a method ofprocessing an object to be decontaminated. The method can compriseprocessing as a single batch in a decontamination process an objecthaving an unknown amount of biological soil disposed thereon and/ortherein and a monitoring device comprising container, a test portionthat includes a predetermined quantity of tracer analyte, and adetection reagent according to any of the above embodiments; afterexposing the test portion to the decontamination process, contacting thetest portion with the detection reagent in the container; and using ananalytical instrument to detect a presence or an absence of thedetectable product, wherein the presence of the detectable productindicates a presence of the tracer analyte on the test portion afterexposing the test portion to the washing process. In any embodiment,processing as a single batch in a decontamination process the object andthe monitoring device can comprise processing as a single batch in adecontamination process the object and a plurality of the monitoringdevices. In any embodiment, processing a plurality of monitoring devicescomprises processing a first device at a first location in an automatedwasher or the automated washer-disinfector and processing a seconddevice at a second location in the automated washer or the automatedwasher-disinfector.

In yet another aspect, the present disclosure provides a kit. The kitcan comprise a container comprising a first end with an opening and asecond end opposite the first end and a test element comprising a testportion to which a test composition is releasably adhered. The containercan include a detection reagent disposed therein. The test compositioncomprises a predetermined quantity of tracer analyte. The container isconfigured to receive the test portion and configured to beoperationally coupled to an analytical instrument. The tracer analyteand the detection reagent each are capable of participating in one ormore chemical reaction that results in the formation of a detectableproduct. In any embodiment of the kit, the test element further cancomprise a reservoir with a solvent disposed therein, wherein thereservoir comprises test element comprises a hollow stem and thereservoir comprises a liquid flow regulator capable of placing thereservoir in fluid communication with the hollow stem. In any of theabove embodiments, the kit further can comprise a means to secure a testelement. In any of the above embodiments, the kit further can comprisean article comprising a receiver, the article configured to restrictfluidic accessibility to the test portion. In some embodiments, thearticle can be a lumened object.

In yet another aspect, the present disclosure provides a system. Thesystem can comprise a monitoring device comprising a test element thatincludes a test portion to which a test composition comprising apredetermined quantity of a tracer analyte is releasably adheredaccording to any one of the above embodiments and an analyticalinstrument capable of detecting the detectable product. The container isconfigured to receive the test portion and configured to beoperationally coupled to the analytical instrument. In any embodiment,the system further can comprise a computer capable of receiving datafrom the analytical instrument and a memory capable of storing thereceived data.

In yet another aspect, the present disclosure provides an article. Thearticle can comprise a homogeneous dried composition removably adheredthereto, wherein the composition comprises a predetermined amount ofadenosine-5′-triphosphate. In any embodiment of the article, thecomposition further can comprise a dye in an amount sufficient to beoptically detectable. In any of the above embodiments of the article,the composition further can comprise a polymeric binder.

In yet another aspect, the present disclosure provides a homogeneous,dried artificial test soil consisting essentially ofadenosine-5′-triphosphate, or a salt thereof, and a polymeric binder. Inany embodiment of the test soil, the polymeric binder can comprisepolyvinyl alcohol, polyethylene glycol, or mixtures thereof.

The words “preferred” and “preferably” refer to embodiments of theinvention that may afford certain benefits, under certain circumstances.However, other embodiments may also be preferred, under the same orother circumstances. Furthermore, the recitation of one or morepreferred embodiments does not imply that other embodiments are notuseful, and is not intended to exclude other embodiments from the scopeof the invention.

The terms “comprises” and variations thereof do not have a limitingmeaning where these terms appear in the description and claims.

As used herein, “a,” “an,” “the,” “at least one,” and “one or more” areused interchangeably. Thus, for example, “a” test element can beinterpreted to mean “one or more” test elements.

The term “and/or” means one or all of the listed elements or acombination of any two or more of the listed elements.

Also herein, the recitations of numerical ranges by endpoints includeall numbers subsumed within that range (e.g., 1 to 5 includes 1, 1.5, 2,2.75, 3, 3.80, 4, 5, etc.).

The above summary of the present invention is not intended to describeeach disclosed embodiment or every implementation of the presentinvention. The description that follows more particularly exemplifiesillustrative embodiments. In several places throughout the application,guidance is provided through lists of examples, which examples can beused in various combinations. In each instance, the recited list servesonly as a representative group and should not be interpreted as anexclusive list.

Additional details of these and other embodiments are set forth in theaccompanying drawings and the description below. Other features, objectsand advantages will become apparent from the description and drawings,and from the claims.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is an exploded view of one embodiment of a monitoring devicecomprising a unitary container, shown in cross-section, and a testelement according to the present disclosure.

FIG. 2 is a side view of a test element, partially in section, having analternative test portion comprising a foam material.

FIG. 3 is a side view of a test element, partially in section, having analternative test portion comprising a fibrous material.

FIG. 4 is a side view of a test element, partially in section, having analternative test portion comprising a plurality of recessed areas.

FIG. 4A is a detail view of the test portion of the test element of FIG.4.

FIG. 5A-C are side views, partially in section, of a portion of oneembodiment of an alternative test element comprising a hollow stem, adeformable reservoir, and a breakable valve that places the reservoir inselective fluid communication with the stem, showing how deformation ofthe reservoir causes breakage of the valve permitting the flow of aliquid into the stem.

FIG. 6 is a block diagram of one embodiment of a method of assessing theefficacy of a washing process according to the present disclosure.

FIG. 7 is a side view, partially in section of the assembled monitoringdevice of FIG. 1 with the test element disposed in a first operationalposition with respect to the container.

FIG. 8 is a side view, partially in section of the assembled monitoringdevice of FIG. 1 with the test element disposed in a second operationalposition with respect to the container.

FIG. 9 is a schematic view of a system for assessing the efficacy of awashing process according to the present disclosure.

FIG. 10 is a side view of the handle portion of one embodiment of amonitoring device comprising a secural element according to the presentdisclosure.

FIG. 11 is a perspective view of one embodiment of a process challengedevice, in an open configuration with a test element disposed therein,according to the present disclosure

DETAILED DESCRIPTION

Before any embodiments of the present disclosure are explained indetail, it is to be understood that the invention is not limited in itsapplication to the details of construction and the arrangement ofcomponents set forth in the following description or illustrated in thefollowing drawings. The invention is capable of other embodiments and ofbeing practiced or of being carried out in various ways. Also, it is tobe understood that the phraseology and terminology used herein is forthe purpose of description and should not be regarded as limiting. Theuse of “including,” “comprising,” or “having” and variations thereofherein is meant to encompass the items listed thereafter and equivalentsthereof as well as additional items. Unless specified or limitedotherwise, the terms “connected” and “coupled” and variations thereofare used broadly and encompass both direct and indirect connections andcouplings. Further, “connected” and “coupled” are not restricted tophysical or mechanical connections or couplings. It is to be understoodthat other embodiments may be utilized and structural or logical changesmay be made without departing from the scope of the present disclosure.Furthermore, terms such as “front,” “rear,” “top,” “bottom,” and thelike are only used to describe elements as they relate to one another,but are in no way meant to recite specific orientations of theapparatus, to indicate or imply necessary or required orientations ofthe apparatus, or to specify how the invention described herein will beused, mounted, displayed, or positioned in use.

The present disclosure relates to a process equipment monitoring deviceand a method of use thereof. The process equipment monitoring device canbe used to test the efficacy of a washing process and, in particular, awashing process conducted by an automated washer. The monitoring deviceis an adaptation of any one of a variety of existing test devices thatare currently used to detect the presence or quantity of a chemicalanalyte. Each of the existing monitoring devices comprises a sampleacquisition device (e.g., a swab or dipstick) that is configured forcontact with a liquid or solid sample such that the sample acquisitiondevice retains at least a portion of the sample. The sample acquisitiondevice is subsequently inserted into a container where it contacts adetection reagent that reacts directly or indirectly with the analyte toform a detectable product (i.e., a colored compound) that can beobserved, and optionally quantitated, by visible inspection or by usingan analytical instrument (e.g., a spectrophotometer or luminometer).

The inventive process equipment monitor includes a test element, whichis analogous to the sample acquisition device described above, but hasbeen modified in a way that renders it substantially unsuitable for itsoriginal purpose (i.e., to detect unknown quantities of analyte). Themodification includes deliberately adulterating the test element with apredetermined quantity of tracer analyte that the existing test devicewas designed to detect. Furthermore, tracer analyte is applied to thetest element in such a way that a portion or all of the tracer analyteis released from the test element when the test element is exposed to awashing process that meets or exceeds predefined standards for efficacy.

The above-mentioned existing test devices are used to detect thepresence, absence, or quantity of a variety of analytes including, forexample, biological analytes such as carbohydrates (e.g., glucose),protein, nucleic acid, and hemoglobin. Typically, the test devicesinclude a sample-acquisition component (e.g., a swab, a pipette, asponge, or the like) to obtain a sample to be tested and a containerinto which the sample acquisition device and/or sample can be placed inorder to detect the analyte. The container may include a detectionreagent disposed therein, the detection reagent capable of interactingwith the analyte to form a detectable moiety (e.g., a chemicalderivative of the analyte or a detectable byproduct of the interactionsuch as light, for example). In addition, many of the test devices areadapted to be used with an analytical instrument to obtain the result ofthe test. For example, the container of the test device may be shapedand dimensioned so that at least a portion of the container can beinserted into the analytical instrument and the result is automaticallyread, and optionally exported and/or electronically saved, by theinstrument.

Nonlimiting examples of such existing test devices include the 3MCLEAN-TRACE Surface ATP Swab available from 3M Company (St. Paul,Minn.), the AQUASNAP ATP Water Test available from Hygiena (Camarillo,Calif.), the ACCUPOINT 2 ATP Sanitation Monitoring System available fromNeogen Corporation (Lansing, Mich.), and the PRO-CLEAN Rapid ProteinResidue Test available from Hygiena.

The inventive monitoring devices of the present disclosure embody atleast one modification of these test devices. The monitoring devices aremodified such that the sample acquisition component of the original testdevice (hereinafter, called the “test element” of the modifiedmonitoring device) is adulterated with a test composition that comprisesa predetermined quantity of the tracer analyte the test device isdesigned to detect. In contrast to a typical prior art test device,which is configured to detect the absence, presence, or quantity of aparticular analyte; the modified test device (i.e., the monitoringdevice) is configured to detect whether the test element has beenexposed to an environment that diminished or eliminated the traceranalyte-containing test composition imbued thereon.

In one aspect, the present disclosure provides a monitoring device. Themonitoring device can be used in a variety of methods disclosed herein.FIG. 1 shows an exploded view of one embodiment of a monitoring device100 according to the present disclosure. The monitoring device 100comprises a container 10 and a test element 40. The test element 40comprises a test portion 44, to which a test composition 50 isreleasably adhered, and a handle 49. The container 10 has a first end 12and a second end 16 opposite the first end. The first end 12 ofcontainer 10 comprises opening 14 into which at least a portion of atest element 40 can be inserted.

The container 10 can be formed (e.g., by injection molding or extrusion)of polymeric materials (e.g., polyethylene, polypropylene) as a unitarypart. As with the existing test devices described herein, when detectionof the tracer analyte comprises optical detection of a product derivedtherefrom, the container 10 should be formed using materials andprocesses that permit the transmission of wavelengths of light that aresuitable to permit optical detection of the product.

Optionally, the monitoring device 100 further may comprise a frangibleseal 35 disposed in the container 10. The frangible seal 35, if presentcan partition the container 10 into two chambers, a receiving chamber 19proximate the opening 14 and a cuvette chamber 22 distal the opening 14.The frangible seal 35 can be made from a water-resistant material suchas, for example, a plastic film, a metal foil, or a metal-coated plasticfilm. The frangible seal 35 can be coupled to the container 10 viacoupling means that are known in the art (e.g., an adhesive, anultrasonic weld, and the like). The frangible seal 35 may be directlycoupled (not shown) to the container 10 at a structure such as flange23, for example. Alternatively, the frangible seal 35 can be coupled(e.g., via an adhesive, an ultrasonic weld, or the like) to a separatestructure (e.g., sealing member 32), which can be inserted into thecontainer 10 and disposed against flange 23, as shown in FIG. 1. Thesealing member 32 can be formed from a relatively flexible and/ormalleable material such as, for example, polyethylene, polypropylene,silicone, or butyl rubber. Preferably, the frangible seal 35 and sealingmember 32, if present, form a liquid-resistant barrier between thereceiving chamber 19 and the cuvette chamber 22.

The container 10 can be formed (e.g., by injection molding or extrusion)of polymeric materials (e.g., polyethylene, polypropylene, polystyrene,polycarbonate). The walls of the cuvette portion 20 can be molded, forexample, to form one of a variety of geometric shapes such as, forexample, cubic, cuboid, cylindrical, conical, frusto-conical, othergeometric shapes suitable to be operationally coupled to an analyticalinstrument (not shown). Preferably, the wall 24 of the cuvette portion20 can be configured (e.g., by using a relatively transparent ortranslucent material and/or by constructing the cuvette portion with atleast one relatively thin wall 24) to permit the transmission of light(e.g., visible light) into and/or out of the cuvette portion.

An optional lamina 70 can be affixed (e.g., adhesively affixed) to thecontainer (e.g., proximate the opening) The lamina 70 can be made frompaper or a plastic film, for example, and may be used as a label.

The monitoring device further comprises a detection reagent 55 disposedin the container. In the illustrated embodiment, the detection reagent55 is disposed in the container as a solid (e.g., a solid powder). Inany embodiment, the detection reagent may be dissolved or suspended in asolvent as described below. In some embodiments (not shown), themonitoring device may comprise a second frangible seal disposed betweenthe first frangible seal and the opening. The space between the firstand second frangible seals forms a compartment in which the reagent,either in dry (e.g., powder) or liquid form, can be disposed.

Optionally, in any embodiment, the container can include a solventdisposed therein. In the illustrated embodiment, the first solvent 60 isdisposed in the cuvette chamber 22. In any embodiment (not shown), thesolvent alternatively or additionally may be disposed in the receivingchamber 19. In any embodiment, the frangible seal 35 can preventunintended movement of the first solvent 60 between the receivingchamber 15 and the cuvette chamber 22.

In any embodiment, the first solvent 60 can be a liquid in which aportion (e.g., the tracer analyte) or all of the test composition 50 issoluble. In any embodiment, the first solvent 60 may comprise water. Insome embodiments, the first solvent 60 additionally comprises a buffercomponent to maintain the solvent within a predefined pH range (e.g., apH range that is suitable for a reaction used in the detection of thetracer analyte). In some embodiments, the solvent may comprise asurfactant (e.g., a nonionic surfactant) to facilitate the dispersion ofthe tracer analyte and/or test composition 50 into the first solvent 60.A suitable surfactant does not substantially interfere with a reaction,a detection reagent, and/or an instrument that is used for the detectionof the tracer analyte.

A monitoring device of the present disclosure comprises a detectionreagent for detecting the tracer analyte. In some embodiments, themonitoring device may comprise a plurality of detection reagents. Atleast one detection reagent may be disposed in the container. In anyembodiment, at least one detection reagent may be disposed in a sealedchamber (e.g., the cuvette chamber) of the container. In any embodiment,the at least one detection reagent may be dissolved in the solvent. Insome embodiments, (not shown) the detection reagent may be disposed on(e.g., as a coating such as a dried coating) and/or in the test element(e.g., dissolved in a solvent disposed in a reservoir, as disclosedherein). The particular detection reagent disposed in the monitoringdevice is selected according to the tracer analyte and/or the instrumentthat is used to detect the tracer analyte, the derivative of the traceranalyte, or the byproduct of the tracer analyte. A person havingordinary skill in the art will recognize a suitable detection reagentfor a particular tracer analyte. By way of example, suitable detectionreagents to detect a protein tracer analyte include a Cu²⁺ compound(e.g., CuSO₄), sodium tartrate, sodium carbonate, sodium bicarbonate,and bicinchoninic acid. One or more of the foregoing reagents can beprovided in a container according to the present disclosure. By way ofanother example, suitable detection reagents to detect ATP traceranalyte include luciferin and luciferase. In any embodiment, a firstdetection reagent may be provided in one chamber of the container and asecond detection reagent may be provided in another chamber of thecontainer.

In some embodiments, the solvent may comprise a stabilizer (e.g. enzymestabilizers).

Referring back to FIG. 1, the test element 40 comprises a test portion44 and an optional stem 45. The stem 45 can be constructed from avariety of materials, such as wood, plastic, metal, or combinationsthereof. In some embodiments, the stem 45 can be fabricated from asufficiently flexible material (e.g., metal wire or plastic polymer) toinsert the test portion 44 into tortuous spaces. Advantageously, inthose embodiments, the test element can be used to assess the ability ofa washing process to penetrate effectively into the tortuous spaces. Inother embodiments, the stem 45 can be relatively inflexible. The stem 45is adapted to be coupled (e.g., by friction fit or via an adhesive) tothe handle 49. In use, the stem 45 or the handle 49 can be grasped by anoperator in order to avoid contact between the operator and the testportion 44 and or test composition 50.

In any embodiment, the test portion 44 can be a substantially smoothsurface such as, for example, a portion of the stem 45, as illustratedin FIG. 1. Alternatively, the test portion may include additional (e.g.,3-dimensional) structural features. The additional structural featuresprovide a greater challenge to a washing process because the structuralfeatures provide physical obstacles that hinder the removal of the testcomposition 50 from the test portion. In any embodiment, the testcomposition 50 can be applied as a liquid mixture and/or liquidsuspension to the test portion 44 using processes that are known in theart including, for example, kiss coating, dip coating and spray coating.A portion or all of the liquid can subsequently be removed from thecomposition by evaporation (e.g., by placing the test element into abiosafety hood at ambient temperature (e.g., about 23° C.) for about 2-3hours, for example). In the illustrated embodiment of FIG. 1, the testportion 44 is shown in partial section in order to show the testcomposition 50 coated on one side of the test portion and the underlyingstructure (e.g., stem 45) on the other side of the test portion. In anyembodiment, the test composition 50 may be coated on the entirecircumference of the test portion.

FIG. 2 shows one embodiment of a test element 41 having a test portion44 with 3-dimensional structural features. The test element 41 can beused in any embodiment of the monitoring devices, methods, and systemsof the present disclosure. The test portion 44 comprises a foam material46 that is imbued with the test composition 50. The foam material 46comprises individual cells or void spaces in which and to which the testcomposition 50 can be releasably adhered. Suitable foam materials foruse in a test portion 44 of the present disclosure should releasablyretain the test composition 50 thereon and, in particular, shouldreleasably retain the tracer analyte. Non-limiting examples of suitablefoam materials include polyurethane foams, polyethylene foams, andpolystyrene foams. In some embodiments, the foams may be treated (e.g.,corona-treated or electron beam-treated) in order to make the surface ofthe polymer more hydrophilic. The foam material 46 can be coupled to thestem 45, if present, or handle 49 using materials (e.g., melt bond,ultrasonic weld, adhesives, mechanical fasteners, or the like) andprocesses known in the art. In the illustrated embodiment of FIG. 2, thetest portion 44 is shown in partial section in order to show the testcomposition 50 coated on one side of the test portion and the underlyingstructure (e.g., foam material 46) on the other side of the testportion. In any embodiment, the test composition 50 may be coated on theentire circumference of the test portion.

FIG. 3 shows an alternative embodiment of a test element 42 having atest portion 44 with 3-dimensional structural features. The test element42 can be used in any embodiment of the monitoring devices, methods, andsystems of the present disclosure. The test portion 44 comprises afibrous material 47. The fibrous material 47 may comprise nonwovenfibers, as shown in FIG. 3, or woven fibers (not shown). The fibrousmaterial comprises individual fibers with void spaces there between. Thetest composition can be releasably adhered to the surface of the fibersand, optionally, may fill void spaces between the fibers. Suitablefibrous materials for use in a test portion 44 of the present disclosureshould releasably retain the test composition 50 thereon and, inparticular, should releasably retain the tracer analyte. Non-limitingexamples of suitable fibrous materials include cotton, DACRON polyester,rayon, nylon, flocked nylon, polyester, polypropylene, polyethylene. Insome embodiments, the fibrous material may be treated (e.g.,corona-treated, electron beam-treated, or coated with diamond-likeglass) in order to make the surface of the material more hydrophilic.The fibrous material can be coupled to the stem 45, if present, orhandle 49 using materials (e.g., adhesives, mechanical fasteners, or thelike) and processes (e.g., fiber entanglement) known in the art. In theillustrated embodiment of FIG. 3, the test portion 44 is shown inpartial section in order to show the test composition 50 coated on oneside of the test portion and the underlying structure (e.g., fibrousmaterial 47) on the other side of the test portion. In any embodiment,the test composition 50 may be coated on the entire circumference of thetest portion.

FIG. 4 shows another alternative embodiment of a test element 43 havinga test portion 44 with 3-dimentional structural features. The testelement 43 can be used in any embodiment of the monitoring devices,methods, and systems of the present disclosure. The test portion 44 inthis embodiment comprises one or more cavity 48. In this embodiment, thestem 45 and test portion 44 may be formed as a unitary part or may beformed as separate parts that are coupled together (e.g., by frictionfit or via an adhesive). The test portion 44 may be formed at least inpart of relatively rigid polymer (e.g., nylon, polysulfone,polycarbonate, or combinations thereof) or it may be formed using a morecompliant polymer, such as silicone. Suitable materials for test portion44 include, but are not limited to, any thermoplastic materials suitablefor casting, profile extrusion, molding (e.g., injection molding) orembossing including, for example, polyolefins, polyesters, polyamides,poly(vinyl chloride), polymethyl methacrylate, polycarbonate, nylon, andthe like. In other embodiments, test portion 44 may be formed by moldingor embossing a sheet of suitable material into the desired cavitystructure. In some embodiments, the test portion 44 may be treated(e.g., corona-treated or electron beam-treated) in order to make thesurface of the material more hydrophilic. In the illustrated embodimentof FIG. 4, the test portion 44 is shown in partial section in order toshow the test composition 50 coated on one side of the test portion andthe underlying structure (e.g., cavities 48) on the other side of thetest portion. In any embodiment, the test composition 50 may be coatedon the entire circumference of the test portion.

In any embodiment, the test portion can comprise a porous fibrousnonwoven matrix (not shown). In any embodiment, the nonwoven matrix cancomprise a wet-laid fiber matrix fabricated from polyethylene fibers(e.g., 1 denier fibrillated polyethylene fibers), nylon fibers (e.g., 6denier, 5.08-cm chopped nylon fibers), bicomponent polymeric fibers(e.g., 1 denier bicomponent ethylene vinyl acetate/polypropylenefibers), or glass fibers. In any embodiment, the tracer analyte (e.g., aplurality of viable microorganisms as described herein) can be disposedon and/or in the fibrous nonwoven matrix. Processes for the productionof suitable wet-laid fiber matrixes are described, for example inInternational Publication No. WO 2012/078426, which is incorporatedherein by reference in its entirety.

In any embodiment, the test portion further comprises a plurality ofinorganic particles such as the concentration agent particles (e.g.,amorphous silicates of metals such as magnesium, calcium, zinc,aluminum, iron, titanium, and the like, and combinations thereof)described in International Patent Publication No. WO 2009/085357, whichis incorporated herein by reference in its entirety. In any embodiment,the plurality of inorganic particles can be dispersed in the fibrousnonwoven matrix described herein. In any embodiment, when the traceranalyte comprises a plurality of viable microorganisms, the plurality ofmicroorganisms can be adhered (e.g., releasably adhered) to two or moreof the plurality of inorganic particles. International PatentPublication No. WO 2009/085357 discloses processes for binding aplurality of viable microorganisms to the inorganic particles of thepresent disclosure.

A person having ordinary skill in the art will recognize a variety ofdesign configurations can be used for the one or more cavity in the testelement 43. For example, International Publication No. WO 2009/134509,which is incorporated herein by reference in its entirety, discloses avariety of sample acquisition devices comprising cavities that aresuitable for use as in a test portion 44 of a test element 43.International Patent Publication No. WO 1993/00994, which isincorporated herein by reference in its entirety, also discloses asample acquisition device with a plurality of grooves capable ofretaining a sample. One or more of the grooves described therein couldbe used in a test element according to the present disclosure.

In any embodiment, the test element may be configured to actuate (i.e.,open) the frangible seal. Referring back to FIG. 4, the test element 43comprises a piercing tip 52 that is shaped to puncture a frangible seal.Alternatively or additionally, the stem 45 of any test element can beformed from a material (e.g., wood, metal, plastic) that is rigid enoughsuch that, when urged against a frangible seal, the stem can deformand/or rupture the frangible seal.

In any embodiment, the test portion of the test element can be shapedlike a medical instrument or a part thereof. Advantageously, when themonitoring device of this embodiment is used to assess the efficacy of awashing process, the washing process is challenged to remove material(i.e., the test composition) from an object that may be similar to theactual medical instruments that are cleaned in the automated washer. Insome embodiments, the test portion may comprise a hinge structure (e.g.,a hinge structure found on a scissors or a medical clamp).Advantageously, in these embodiments, the removal of the testcomposition from the test portion more closely resembles actualconditions in a cleaning process.

The test composition is releasably adhered to the test portion of thetest element. The test composition is dispersible, and may be soluble,in an aqueous solvent (e.g. an aqueous solvent used to was articles).The test composition comprises a tracer analyte. The tracer analyte isdispersible, and may be soluble, in an aqueous solvent (e.g. an aqueoussolvent used to wash articles). A “tracer analyte”, as used hereincomprises a compound that can be quantitatively detected using aphoto-optical device. The detection may be achieved by direct detection(e.g., using an optical property of the tracer analyte per se such as,for example the U.V-visible absorbance of the tracer analyte) or byindirect detection (e.g., using an optical property of a derivative orbyproduct of the tracer analyte). In any embodiment, the tracer analytecan be a chemical compound that is capable of participating in achemical reaction that, either directly or indirectly, results in adetectable product. “Chemical reaction”, as used herein, includesbinding reactions (e.g., ionic binding, covalent binding, or hydrophobicinteraction), synthetic reactions, decomposition reactions, oxidationreactions, reduction reactions, complexation reactions, acid-basereactions, and photochemical reactions.

By way of example, in one embodiment, the tracer analyte comprises anunlabeled protein (e.g., bovine serum albumin). In this embodiment, thetracer analyte can be detected indirectly by reacting the tracer analytewith a protein-detecting detection reagent such as bicinchoninic acid,for example, thereby forming a byproduct (i.e., a purple-coloredbicinchoninic acid-Cu¹⁺ chelate), which can be quantitated using aspectrophotometer device. By way of example, in another embodiment, thetracer analyte can comprise a labeled protein that undergoes a chemicalreaction (e.g., a binding reaction, a hydrolytic reaction) to bind,release, or detectably modify the label and/or the labeled protein. Byway of example, in yet another embodiment, the tracer analyte comprisesadenosine-5′-triphosphate (ATP) or a molecule (e.g., ADP) that can beconverted to ATP. In this embodiment, the ATP can be quantitativelydetected, for example, by reacting it with luciferin and luciferase tocause the emission of a byproduct (light), which can be detectedquantitatively using a luminometer. A person having ordinary skill inthe art will recognize other compounds that are suitable for use as atracer analyte and the particular detection reagent(s) and/orinstrument(s) that can be used to detect and quantitate the traceranalyte.

In any embodiment, the tracer analyte can be selected from the groupconsisting of a plurality of viable microorganisms or a biomoleculeassociated therewith, an acid, a base, a nucleotide, a protein, anucleic acid, a carbohydrate, or hemoglobin. The acid may comprise anorganic acid (e.g., a fatty acid). The base may comprise an organic base(e.g., a basic amino acid such as arginine or lysine). The acid or basetracer analyte may be detected by U.V-visible absorbance or apH-detecting detection reagent (e.g., a pH indicator) and quantitatingthe acid or base using a spectrophotometer, for example.

As discussed above, in any embodiment, the tracer element can comprise aplurality of viable microorganisms. Preferably, the viablemicroorganisms are generally regarded as non-pathogenic microorganisms.Suitable non-pathogenic microorganisms include, for example bacteria(e.g., probiotic bacteria such as Lactobacillus and Bifidobacteriumspecies) and yeast (e.g., Saccharomyces cerevisiae), which can bedetected by culture methods or by detecting biomolecules (e.g.,proteins, nucleic acids, small molecules (e.g., ATP), and/or antigens)associated therewith. Suitable nonpathogenic microorganisms also includebacterial endospores (e.g., spores of Bacillus atrophaeus or Geobacillusstearothermophilus). Coatings comprising about 10⁶ to about 10⁸ sporescan be detected by enzymes (e.g., glucosidases or proteases) associatedtherewith (see, for example, U.S. Pat. No. 5,073,488 and U.S. PatentApplication Publication No. 2011/0182770; which are incorporated hereinby reference in their entirety) or by using nucleic acid-binding dyes(see, for example, U.S. Patent Application Publication No. 2011/0200992,which is incorporated herein by reference in its entirety).

In any embodiment, the test composition further may comprise biologicalmaterials that are found in animal tissue, fluids, and/or excreta.Nonlimiting examples of said biological materials include blood cells,serum, bilirubin, mucin, and carbohydrates.

In any embodiment, the test composition further may comprise a dye thatis visually detectable prior to exposing the test element to a washingprocess. Accordingly, the dye can permit visual confirmation that thetest element has the test composition coated thereon.

In any embodiment, the test composition optionally may comprise apolymeric binder. Advantageously, the polymeric binder inhibits thedispersion of the tracer analyte in an aqueous solvent. Without beingbound by theory, this inhibition occurs because the polymeric binderacts as a diffusion barrier to inhibit the release of the tracer elementfrom the test portion. The polymeric binder rehydrates and dissolvesinto an aqueous washing solvent relatively slowly compared to the traceranalyte. In any embodiment, the polymeric binder may comprise polyvinylalcohol (PVA). The polyvinyl alcohol can be prepared as an aqueoussolution comprising the tracer analyte (e.g., 1 microgram/mL ATP), whichis coated onto the test element and dried, as described herein. In anyembodiment, the PVA-containing aqueous solution may comprise about 9weight percent to about 11 weight percent PVA. In any embodiment, thepolymeric binder may comprise polyethylene glycol (PEG 8000, which has amolecular weight of approximately 8,000 daltons). The polyethyleneglycol can be prepared as an aqueous solution comprising the traceranalyte (e.g., 1 microgram/mL ATP), which is coated onto the testelement and dried, as described herein. In any embodiment, thePEG-containing aqueous solution may comprise about 50 weight percentPEG.

In any embodiment, the test composition may be prepared as a homogeneousmixture in a suitable solvent (e.g., water and/or an alcohol). In anyembodiment, the test composition may be dissolved or suspended in anorganic solvent before it is applied to the test element.Advantageously, this may permit the application of higher concentratedsolutions of tracer analyte (or other components of the testcomposition) wherein the tracer analyte and/or component is dissolved ata concentration that exceeds the water solubility of the tracer analyteor component.

In any embodiment, the test composition can be applied as a singlesolution and/or suspension to the test portion of the test element(e.g., using processes described herein) in a single application.Alternatively, the test composition can be applied to the test portionof the test element as two or more separate solutions and/orsuspensions. For example, a first solution and/or suspension comprisingthe tracer analyte may be applied to the test element and a secondsolution and/or suspension comprising a polymeric binder may be appliedseparately to the test element. Optionally, the first solution and/orsuspension may be permitted to dry or partially dry before the secondsolution and/or suspension is applied.

In any embodiment, the monitoring device can comprise a test elementthat is adapted to deliver a liquid to the container. Nason (U.S. Pat.No. 5,266,266; which is incorporated herein by reference in itsentirety) discloses a specimen test unit that includes a swab memberthat can be adapted to function as a test element according to thepresent disclosure. FIGS. 5A-C show a portion (i.e., the portionproximate the first end 112 of the container 110) of one embodiment of atest element 140 that is adapted to deliver a second solvent 175 to thecontainer 110. In this embodiment, the handle 149 comprises a hollowchannel 162 extending there through. Coupled to the handle 149 (e.g.,via an adhesive (not shown) or by friction fit) is a reservoir 164 witha hollow stem 145 coupled thereto (e.g. by friction-fit).

A portion 180 of the hollow stem 145 disposed in the reservoir 164comprises a liquid flow regulator (e.g., a breakable liquid flowregulator) capable of placing the reservoir in fluid communication withthe hollow stem 145. The portion 180 includes a solid rod segment 182and a score 184 that facilitates the breakage of the stem 145, therebycreating a stem opening 186 to permit liquid flow through out of thereservoir through the hollow stem 145. The test element 140 can be madeas described by Nason. As shown in FIG. 5B, (e.g., manual pressure)pressure against the flexible reservoir 164 in the direction of arrow“A” causes the reservoir 164 to deflect against the rod segment 182,causing the score 184 to fracture and optionally separate from thehollow stem 145 (as shown in FIG. 5C), which permits the flow of secondsolvent 175 through the stem opening 186 and into the hollow stem 145,as shown by arrow “B”. A person having ordinary skill in the art willrecognize other liquid flow regulator means (e.g., frangible ampoulesand other means disclosed in U.S. Pat. Nos. 4,978,504 and 5,879,635,which are incorporated herein by reference in their entirety) that canbe used to place the second solvent 175 in the reservoir 164 into fluidcommunication with the hollow stem 145.

In any embodiment, the second solvent 175 can be a liquid in which aportion (e.g., the tracer analyte) or all of the test composition (notshown) is soluble. In any embodiment, the second solvent 175 maycomprise water. In some embodiments, the second solvent 175 additionallycomprises a buffer component to maintain the solvent within a predefinedpH range (e.g., a pH range that is suitable for a reaction used in thedetection of the tracer analyte). In some embodiments, the solvent maycomprise a surfactant (e.g., a nonionic surfactant) to facilitate thedispersion of the tracer analyte and/or test composition 50 into thesecond solvent 175. A suitable surfactant does not substantiallyinterfere with a reaction, a detection reagent, and/or instrument thatis used for the detection of the tracer analyte. In any embodiment thesecond solvent 175 may be the same as the first solvent (not shown), ifpresent in the monitoring device.

In another aspect, the present disclosure provides a first method. Forexample, the present disclosure provides a first method of assessing theefficacy of a washing process. FIG. 6 shows a block diagram of oneembodiment of a first method of assessing the efficacy of a washingprocess according to the present disclosure. The first method 1000comprises the step 90 of exposing to the washing process the testportion of any embodiment of a monitoring device according to thepresent disclosure, the step 92 of contacting the test portion of thetest element with the detection reagent in the container of themonitoring device, and the step 94 of using an analytical instrument todetect a presence or an absence of the detectable product resulting fromthe one or more chemical reaction in which the detection reagent and thetracer analyte are capable of participating.

Exposing the test portion of the test element to the washing process cancomprise placing the test element into an automated washer. In anyembodiment, the automated washer can comprise an automated washerdisinfect such as a GETINGE 46-series washer disinfector (available fromGetinge USA, Inc., Rochester, N.Y.), for example. During normal handlingand use, the test element typically is grasped and/or secured preferablyusing its handle, if present, or its stem. In any embodiment, the testelement can be placed in a rack, which is placed in the automated washerprior to exposing the test element to the washing process. Optionally,the test element can be secured to the rack or to a structure (e.g., arack or shelf) in the automated washer.

In any embodiment, the test element further may comprise a securalstructure configured to detachably secure the test element to astructure (e.g., a rack or shelf) in the automated washer. FIG. 10 showsa portion (i.e., the portion proximate the handle) of one embodiment oftest element 40′ comprising a secural element 85. In some embodiments,the secural element 85 may be formed (e.g., by a molding process) of thesame material as the handle 49. The secural element 85 can be shaped anddimensioned to include one or more engagement structures 86 that canreleasably hold a portion of an automated washer rack or wire basket,for example, and thereby hold the monitoring device at a fixed locationwithin an automated washer. In some embodiments, the secural element 85can be formed separately and attached to the test element 40′ using anattachment means known in the art (e.g., an adhesive, a thermal bond, anultrasonic weld, a screw, a rivet, or the like). The secural element canbe fabricated from any material that is not substantially degraded by awashing process. Non-limiting examples of suitable materials includemetal and polymeric (e.g., polypropylene, polyethylene) materials.

Securing the test element may be performed using a zip-tie, a clamp(e.g., a hose clamp), or the like. In any embodiment, the test portionmay be placed in the automated washer at a peripheral location withinthe washing chamber, thereby testing the efficacy of the washing processin a difficult-to-reach location.

Many commercial automated washers are programmable and are configuredwith preset washing cycles. Accordingly, exposing the test portion ofthe test element to the washing process can comprise placing the testportion into an automated washer and performing at least a portion of anautomated wash cycle while the test portion is disposed in the automatedwasher. An automated washing cycle may comprise, for example, one ormore pre-rinse step, one or more wash step, one or more rinse step, oneor more drying step, or a combination of any two or more of theforegoing steps. After exposing the test portion to at least a portionof the automated washing cycle, the amount of tracer analyte remainingon the test element can be analyzed to determine whether the washingcycle removed any or all of the tracer analyte from the test element,thereby indicating the washing efficacy of the portion of the automatedwashing cycle.

In a preferred embodiment, exposing the test portion of the test elementto the washing process comprises placing the test portion into anautomated washer and performing a complete automated wash cycle whilethe test portion is disposed in the automated washer. A non-limitingexample of a preset automated washing cycle includes the followingsteps: a 1-minute pre-rinse step using cold water, a 5-minute washingstep using hot (e.g., 60° C.) water mixed with an enzyme detergent(e.g., a multi-enzyme detergent), two 1-minute rinse steps with hotwater, a 1-minute disinfection step with very-hot (e.g., 90° C.)deionized water, and a 10-minute drying step. Thus, after exposing thetest portion to the complete automated washing cycle, the amount oftracer analyte remaining on the test element can be analyzed todetermine whether the washing cycle removed any or all of the traceranalyte from the test element.

Typically, while the test portion is exposed to the washing process, thecontainer is kept in a location outside the automated washer. After thetest portion has been exposed to the washing process, the test elementcan be removed (e.g., from the automated washer) and inserted into thereceiving chamber of the container. FIG. 7 shows a side view, partiallyin section of one embodiment of a monitoring device 100 with the testelement inserted into the container. In the illustrated embodiment, thetest element 40 is disposed in a first operational position with respectto the container 10. In the first operational position, a first portionof the test element (e.g., the test portion 44 and stem 45) are disposedin the receiving chamber 19 of the container 10 and a second portion ofthe test element (e.g., the handle 49) is operationally coupled (e.g.,by friction fit) with the container 10 proximate the opening 14 of thecontainer.

The first method of the present disclosure comprises contacting the testportion of the test element with the detection reagent in the containerof the monitoring device. In the illustrated embodiment of FIGS. 7-8,this comprises moving (e.g., by applying manual pressure to the handlein the direction of arrow “5”) the test element 40 into a secondoperational position with respect to the container 10, as shown in FIG.8. In the second operational position, the test element 40 has piercedthe frangible seal 35 and the test portion 44 is contacting the firstsolvent 60, in which the detection reagent (not shown) is dissolved. Byway of example, the tracer analyte can be ATP and the first solvent 60may be an aqueous solution with a pH that is suitable to facilitate thereaction of a detection reagent (e.g., luciferase enzyme) with luciferinand the tracer analyte (ATP).

In any embodiment, contacting the test portion of the test element withthe solvent in the container of the monitoring device can furthercomprise dissolving the tracer analyte and/or test composition into thesolvent.

Referring back to FIG. 6, the first method 1000 of the presentdisclosure comprises the step 94 of using an analytical instrument todetect a presence or an absence of the detectable product resulting fromthe one or more chemical reaction in which the detection reagent and thetracer analyte are capable of participating. In any embodiment, thedetectable product can be a colored compound (e.g., a bicinchoninicacid-Cu+ chelate formed by the reaction of protein with Cu2+ in thepresence of bicinchoninic acid) having a detectable absorbance spectrum.In these embodiments, the detectable product can be detected using aspectrophotometer, for example. In any embodiment, using an analyticalinstrument to detect a presence or an absence of the detectable productcan comprise inserting at least a portion (e.g., a cuvette portion) ofthe container of the monitoring device into the analytical instrument.In any embodiment, the detectable product can be electromagneticradiation (e.g., visible light, such as the light emitted by thereaction of luciferin and luciferase with ATP, for example) having acertain wavelength (e.g., about 550 nm to about 620 nm).

Optionally, the first method 1000 of the present disclosure further cancomprise the step 96 of using the analytical instrument to measure aquantity of the detectable product. In preferred embodiments, thequantity of detectable product is proportional to the quantity of traceranalyte, if present, on the test element. In any embodiment, themeasured quantity can be a threshold detectable quantity, which simplyindicates the presence or absence of the detectable product. A personhaving ordinary skill in the relevant art will recognize the thresholddetectable quantity represents the lower limit of detection and isdefined by several parameters including, for example, the reactants, thecontainer, and the analytical instrument. In any embodiment, themeasured quantity can be an absolute quantity, which can be determinedby comparing the detectable quantity to a standard or a plurality ofstandards, for example. In any embodiment, the measured quantity can bea relative quantity (e.g., relative light units detected from alight-emitting reaction).

Optionally, the first method 1000 of the present disclosure further cancomprise the step 98 of comparing the measured quantity of thedetectable product to a predefined standard (i.e., a “control value”).The control value can be selected to indicate a quantity (e.g., amaximum quantity) of detectable product associated with an adequatewashing cycle. Thus, in these embodiments; when the washing cycle isadequate to remove a sufficient quantity of soil (e.g., the traceranalyte) from the test element, the quantity of detectable product maybe less than or equal to the control value.

In another embodiment of the first method, the standard may be ameasurable quantity of tracer analyte that is detected from a testelement (e.g., a “control” test element) that has not been exposed to awashing cycle. In this embodiment, an indication of exposure to anadequate washing cycle can be that the washed test element retains apredetermined percentage (e.g., up to about 50%, up to about 40%, up toabout 30%, up to about 25%, up to about 20%, up to about 15%, up toabout 10%, up to about 5%, up to about 2%, up to about 1%) of the traceranalyte that is detectable from the control test element.

In any embodiment, the first method further comprises using at least onemeasured quantity of tracer analyte to define an action limit. Actionlimits for controlling a multi-step decontamination process aredisclosed in International Patent Publication No. WO 2012/112482, whichis incorporated herein by reference in its entirety. In any embodiment,the first method further can comprise comparing a measured quantity oftracer analyte to a predefined action limit.

In another embodiment of the first method, the standard may be anarbitrary value (e.g., relative light units, micrograms of traceranalyte, or the like) that is selected (e.g., by the user or theprovider of the test element) to indicate the efficacy of the washingprocess.

In any embodiment, an operator may desire to keep a record of thedetection of a presence or measurable quantity of tracer analytedetected from a test element exposed to a particular washing process. Insome embodiments, the record may be an electronic record that is storedon a computer-readable medium using electronic data storage processesthat are well-known in the art. The computer-readable medium maycomprise random access memory (RAM) such as synchronous dynamic randomaccess memory (SDRAM), read-only memory (ROM), non-volatile randomaccess memory (NVRAM), electrically erasable programmable read-onlymemory (EEPROM), FLASH memory, magnetic or optical data storage media,and the like.

In any embodiment of the first method, an operator may desire toassociate a first datum (e.g., the record of the detection of a presenceor measurable quantity of tracer analyte detected from a test element)with a second datum (e.g., other information related to the testelement). In any embodiment, the second datum comprises informationselected from the group consisting of a date, a time, a washingapparatus, an operator, an instrument to be washed, and a combination oftwo or more of any of the foregoing test data. Advantageously,associating the first datum with the second datum can allow the operatorto verify that a particular instrument was present in an automatedwasher with a test element that verified the efficacy of the washingprocess to which both the instrument and the test element were exposed.

In any embodiment of the first method, exposing the test portion of atest element to a washing process can comprise exposing the testportions of a plurality of test elements to a washing process.Advantageously, this embodiment can be used to identify certain spatialregions within an automated washer that do not wash objects aseffectively as other spatial regions within the automated washer. Thistype of information can be used by the operator to make decisions suchas preventative maintenance schedules and/or load configurations, forexample, for particular automated washers. In these embodiments, a firstmonitoring device can be positioned at a first predefined location(e.g., upper rack proximate the back of the washer) and a secondmonitoring device can be positioned at a second predefined location(lower rack proximate the front of the washer). After exposing the firstand second monitoring devices to a washing process, the tracer analyteremaining on the test portion of each monitoring device is measured asdescribed herein and each measured quantity can be compared to a controlvalue and/or can be compared to each other. The control value mayindicate the monitoring devices were exposed to an effective washingprocess or it may indicate the monitoring devices were exposed to anineffective washing process. According to this embodiment, the firstmethod can be used to create a 2-dimensional or 3-dimensional map of theinterior of an automated washer, the map showing specific regions of thewasher and the washing efficacy of each region.

In any embodiment, the first method optionally can comprise the step ofplacing the test portion of the monitoring device in a receiverconfigured to restrict fluidic accessibility to the test portion. Insome embodiments, the receiver may comprise a wall that shields the testportion from a direct spray of wash solvent (e.g., water) emitted from anozzle or orifice in the automated washer. Thus, in order for the washsolvent to impinge on the test portion, it must take an indirect path(e.g., by deflection off a wall or other object present in the automatedwasher. In some embodiments, placing the test portion of the monitoringdevice in a receiver configured to restrict fluidic accessibility to thetest portion comprises placing the test portion into an interior spaceof a lumened object. Exemplary lumened objects include, for example,tubes, scopes, and the like.

In yet another aspect, the present disclosure provides a second methodof controlling a decontamination process. The second method comprisesprocessing as a single batch in a decontamination process an objecthaving an unknown amount of biological soil disposed thereon and/ortherein and a monitoring device comprising a predetermined quantity oftracer analyte according to any one of the embodiments of the monitoringdevice of the present disclosure. The second method further comprises,after exposing the test portion to the decontamination process,contacting the test portion with the detection reagent in the container;and using the analytical instrument to detect a presence or an absenceof the detectable product, wherein the presence of the detectableproduct indicates a presence of the tracer analyte on the test portionafter exposing the test portion to the washing process. Thedecontamination process may be a multi-step decontamination processincluding, without limitation, subprocesses such as soaking, wiping,brushing, scrubbing, washing, contact with a disinfecting agent, contactwith a sterilizing agent, a combination of any two or more of theforegoing subprocesses or it may be an single process such as any of theaforementioned subprocesses.

“Processing as a single batch” for the purposes of the presentdisclosure and claims means the object (e.g., a medical instrument thatwas used in a medical or surgical procedure) is placed with themonitoring device of the present disclosure into a single container(e.g., a tub, wash basin, automated washer, or the like) for the purposeof exposing the object and the monitoring device to substantiallysimilar decontamination conditions (e.g., solvent, detergent,temperature, flow rates, and/or rinse volume). In any embodiment of thesecond method, processing as a single batch in a decontamination processthe object and the monitoring device comprises processing as a singlebatch in a decontamination process the object and a plurality of themonitoring devices. In any embodiment of the second method, processing aplurality of monitoring devices can comprise processing a firstmonitoring device at a first location (e.g., a first location in anautomated washer or automated washer-disinfector) and processing asecond monitoring device at a second location that is spaced apart fromthe first location (e.g., a second location in the automated washer orthe automated washer-disinfector).

Contacting the test portion with the detection reagent in the containercomprises contacting the test portion with the detection reagent in thecontainer of the monitoring device of the present disclosure in order toproduce a detectable product, as described herein. A presence of thedetectable product indicates at least a portion of the tracer analytewas not released from the test element while the test portion wasexposed to the decontamination process, as described herein. In anyembodiment of the second method, processing the object and themonitoring device in a decontamination process comprises processing theobject and the monitoring device in an automated washer or an automatedwasher-disinfector.

In any embodiment of the second method, detecting a presence or anabsence of the detectable product further comprises calculating aquantity of tracer analyte remaining on the test portion after exposingthe test portion to the decontamination process. In any embodiment, thesecond method further comprises comparing the quantity of tracer analyteremaining on the test portion of at least one monitoring device afterexposing the test portion to the decontamination process to apredetermined quantity of tracer analyte associated with an actionlimit. “Action limit”, as used herein refers to a threshold amount ofbiological analyte, detected in the residue collected from an articleaccording to a pre-defined sample collection method and a pre-definedanalyte detection method, which indicates a particular sub-process or aseries of sub-processes failed to remove or inactivate an acceptableamount of biological soil from the article. Action limits can be used todetermine what activities (e.g., re-processing the load, equipmentmaintenance), if any, should be taken as a result of inadequate removalof the tracer analyte from the monitoring device.

In any embodiment of the second method, if the quantity of traceranalyte remaining on the test portion of the at least one monitoringdevice after exposing the test portion to the decontamination process isless than or equal to the predetermined quantity (e.g., an “actionlimit”), the method further comprises releasing the object for use(e.g., for use in a subsequent procedure or for use in a firstsubsequent process (e.g. a wrapping step and/or a sterilization step toprepare the object for re-use in a medical or surgical procedure).Conversely, in any embodiment of the second method, if the quantity oftracer analyte remaining on the test portion of the at least onemonitoring device after exposing the test portion to the decontaminationprocess is greater than or equal to the predetermined quantity, themethod further comprises releasing the object for use in a secondsubsequent process (e.g., re-washing, re-processing, additional cleaningtreatments, or the like).

In yet another aspect, the present disclosure provides a kit. The kitcomprises a container as disclosed herein, the container comprising afirst end with an opening and a second end opposite the first end,wherein the container includes a detection reagent disposed therein. Thekit further comprises a test element comprising a test portion to whicha test composition is releasably adhered, wherein the test compositioncomprises a predetermined quantity of tracer analyte. The test elementcomprises any suitable tracer analyte according to the presentdisclosure. The detection reagent comprises any detection reagentsuitable to participate in a chemical reaction to detect, directly orindirectly, the tracer analyte. The container is configured to receivethe test portion and configured to be operationally coupled to ananalytical instrument, as described above. The tracer analyte and thedetection reagent each are capable of participating in one or morechemical reaction that results in the formation of a detectable product,wherein the detectable product indicates a presence and, optionally, aquantity of tracer analyte in the one or more chemical reaction.

In any embodiment of the kit, the tracer analyte is selected from thegroup consisting of a plurality of viable microorganisms or abiomolecule associated therewith, an acid, a base, a nucleotide, aprotein, a nucleic acid, a carbohydrate, or hemoglobin. In anyembodiment, the test portion comprises a fibrous nonwoven matrix, asdescribed hereinabove. In any embodiment wherein the analyte comprises aplurality of viable microorganisms, the plurality of viablemicroorganisms may be disposed on or in the fibrous nonwoven matrix. Inany embodiment of the kit, the test portion further comprises aplurality of inorganic particles dispersed in the fibrous nonwovenmatrix. In these embodiments, the plurality of viable microorganisms canbe releasably adhered to two or more of the plurality of inorganicparticles.

In any embodiment of the kit, a portion of the test element can bedisposed in the container (e.g., in a first operational position, asdescribed herein. In any embodiment, the container further comprises afirst solvent disposed therein (e.g., in the cuvette chamber, asdisclosed herein). In any embodiment of the kit, the test elementfurther comprises a reservoir with a solvent disposed therein, whereinthe test element comprises a hollow stem, as described herein, and thereservoir comprises a liquid flow regulator (e.g., a breakable liquidflow regulator) capable of placing the reservoir in fluid communicationwith the hollow stem, as described herein.

In any embodiment, the kit further may comprise a means to secure a testelement. The means to secure the test element may comprise a clamp, astring, a wire, a zip-tie, or any other suitable means capable ofsecuring the test element to an object (e.g., a medical instrument, ashelf, a wire basket) in an automated washer.

In any embodiment, the kit further may comprise a process challengedevice. A “process challenge device”, as used herein, refers to awater-insoluble container (e.g., an envelope, a tube, an elongated tube,a box) with at least one opening and an inner volume defined by at leastone wall. In any embodiment, the process challenge device may compriseone or more walls with a plurality of openings. The process challengedevice is configured to receive at least part of the test portion of anyembodiment of the test element of the present disclosure. In anyembodiment, the process challenge device may be configured to receivethe entire test portion of any embodiment of the test element of thepresent disclosure. In any embodiment, the process challenge device maybe configured to receive the entire test element of any embodiment ofthe present disclosure.

FIG. 11 shows one embodiment of a process challenge device 3000according to the present disclosure. The process challenge device 3000comprises a water-insoluble container 3001 having at least one wall3003. The process challenge device 3000 comprises a plurality of walls3003 that can be configured to surround an object placed in the processchallenge device 3000. Optionally, one of the walls is hingedly attachedto another wall via a hinge means 3009, thereby permitting the processchallenge device 3000 to be placed into an open configuration (asillustrated in FIG. 11) or in a closed configuration (not shown). In theopen configuration, the process challenge device 3000 can receive a testelement such as test element 43, for example. The test element can beany embodiment of a test element according to the present disclosure. Ina preferred embodiment, the test element 43 is a test element from amonitoring device (not shown) of the present disclosure.

The at least one wall 3003 presents a physical impediment (i.e.,challenge) to hinder the passage of process fluid (e.g., water and/ordetergent) from a washer or washer-disinfector to an object (e.g., atest element) disposed in the process challenge device 3000. In anyembodiment, the severity of the challenge can be attenuated by providingrelatively greater access (i.e., pathways) to the test element. Thus,optionally, at least one wall 3003 of the one or more walls of theprocess challenge device 3000 comprises at least one hole 3005 extendingtherethrough. The at least one hole 3005 can be any size (e.g.,diameter) that is suitable to permit the passage of a fluidtherethrough. In a particular embodiment, the size of the holes 3005 aresimilar, as illustrated in FIG. 11. In another particular embodiment(not shown), the size of the holes 3005 are different. The holes 3005may be about 1 mm to about 10 mm or greater in diameter, for example.

In any embodiment, at least one wall 3003 of the one or more walls ofthe process challenge device 3000 comprises a plurality of holes 3005extending therethrough. In any embodiment, a plurality of walls 3003 ofthe one or more walls of the process challenge device 3000 comprises aplurality of holes 3005 extending therethrough, as shown in FIG. 11. Theholes 3005 permit the passage of process fluid (not shown) therethrough.

In any embodiment, the process challenge device 3000 optionallycomprises means 3007 for securing an object (e.g., test element 43) inthe process challenge device 3000. In the illustrated embodiment of FIG.11, the means 3007 comprises a spring-like structure capable ofentrapping a portion (e.g. handle 49 of test element 43) of the object.A person having ordinary skill in the art will recognize other suitablemeans such as, for example, a clip, a clamp, a hook, a tether, anadhesive, a weighted object, and the like. In any embodiment, the means3007 secures the test element 43 such that the test composition 50 isheld in a generally predetermined position within the process challengedevice 3000.

In use, the process challenge device 3000, with the test element 43disposed therein, can be placed in any suitable location in a washer orwasher-disinfector. The washer or washer-disinfector can be operatedaccording to various operating parameters (e.g., standard operatingparameters) and the test element 43 can be removed subsequently andtested as described herein in order to determine whether the testcomposition 50 was adequately dislodged from the test element during theoperating cycle. Accordingly, in any embodiment of the methods ofassessing the efficacy of a washing process, the method furthercomprises, before exposing the test portion of the monitoring device tothe washing process, positioning at least a part of the test portion ofthe monitoring device in a process challenge device.

Process challenge devices of the present disclosure can be made fromsuitable water-insoluble materials (e.g., stainless steel, aluminum,polymers such as HDPE or polycarbonate, for example) according toconventional processes (e.g., molding) known in the art. In anyembodiment, the material(s) from which the process challenge device ismade can be relatively rigid, relatively flexible, or may contain aportion that is relatively rigid and a portion that is relativelyflexible. In any embodiment, the hinge means 3009, if present, maycomprise a spring hinge or biasing hinge that is adapted to hold thedevice 3000 in a normally-closed configuration.

In another aspect, the present disclosure provides a system. The systemcan be used to test the efficacy of a washing process. The systemcomprises a monitoring device comprising a container and a test elementaccording to any embodiment described herein. The test element comprisesa predetermined quantity of a tracer analyte releasably adhered thereto.The system further comprises an analytical instrument capable ofdetecting a detectable product that indicates a presence and,optionally, a quantity of tracer analyte in one or more chemicalreaction. In some embodiments, the system may further comprise acomputer capable of receiving data from the analytical instrument and amemory (not shown) capable of storing the received data. In anyembodiment, the system further comprises the process challenge devicedisclosed herein. FIG. 9 shows a schematic view of one embodiment of asystem 2000 according to the present disclosure. The system comprises amonitoring device 2100 and an analytical instrument 2500. Optionally,the system 2000 further comprises a computer 2600.

Optionally, the computer 2600 may comprise software or firmware capableof operating the analytical instrument 2500. In any embodiment, thesoftware may be adapted to facilitate the detection of the detectableproduct that indicates a presence of the tracer analyte. The computer2600 may include memory and may create an information database in itsmemory to track and store such information. Computer 2600 may associatevarious types of information with the monitoring device 2100. Numericalvalues associated with one or more test elements may be analyzed and/orstored by computer 2600. In addition, a numerical value associated witha first test element can be analyzed by computer 2600 to compare thevalue to second test element and/or a control value associated with astandard (e.g., a positive control, a negative control).

Computer 2600 may include a microprocessor that executes software foranalysis of monitoring device 2100, and for database managementconsistent with the techniques known in the art. Accordingly, computer2600 may also include memory to store the various types of informationassociated with a particular monitoring device 2100. Computer 2600 maycomprise a personal computer (PC), desktop computer, laptop computer,handheld computer, workstation, or the like.

In another aspect, the present disclosure provides a homogeneous, driedartificial test soil. The test soil is a composition comprising a targetanalyte (e.g., an acid, a base, a nucleotide, a protein, a nucleic acid,a carbohydrate, or hemoglobin) according to the present disclosure. In apreferred embodiment, the target analyte is Adenosine-5′-triphosphate.In any embodiment, the test soil composition further can comprise anoptional dye in an amount sufficient to be optically detectable, asdescribed herein. The test soil further comprises a polymeric binder.The dried test soil is prepared by dissolving and/or making ahomogeneous dispersion of the target analyte, polymeric binder, andoptional dye in a suitable solvent (e.g., water, alcohol, or mixturesthereof), applying the mixture to a surface (e.g., by spraying,dip-coating, or other coating processes known in the art) of asubstrate, and removing at least a portion of the solvent (e.g.,substantially all of the solvent) to obtain a dried coating on thesubstrate.

Without being bound by theory, the polymeric binder provides one or moreof the following technical effects in the test soil composition: 1) thepolymeric binder provides bulk mass that can facilitate the adherence ofrelatively small quantities of tracer analyte to a substrate, 2) thepolymeric binder provides adhesive properties to facilitate theadherence of the test soil composition to the substrate, 3) thepolymeric binder provides a solubility and/or diffusion barrier thatprevents the substantially immediate dissolution and release of thetracer analyte from the substrate when the test soil is contacted with asolvent (e.g., water, hot water) in which it is soluble or dispersibleand, 4) in the instance where the polymeric binder comprises a protein,the polymeric binder may provide some buffering capacity to maintain thepH of the composition.

Suitable polymeric binders include, for example, polyols (e.g.,polyvinyl alcohol, polysaccharides), polyethers (e.g., polyethyleneoxides), and polyamides (e.g., proteins such as serum albumin, forexample). The molecular weight of the polymeric binder can be selectedsuch that the binder is more or less soluble in the liquid used in thewashing process. For a given coating weight, higher molecular weightbinders may be used to produce test soil compositions that are moredifficult to wash off. Conversely, for a given coating weight, lowermolecular weight binders may be used to produce test soil compositionsthat are less difficult to wash off.

EMBODIMENTS

Embodiment A is a monitoring device, comprising:

a test composition comprising a predetermined quantity of traceranalyte;

a test element comprising a test portion to which the test compositionis releasably adhered;

a detection reagent; and

a container comprising a first end with an opening and a second endopposite the first end;

wherein the container is configured to receive the test portion andconfigured to be operationally coupled to an analytical instrument;

wherein the tracer analyte and the detection reagent are capable ofparticipating in one or more chemical reaction that results in theformation of a detectable product.

Embodiment B is the monitoring device of Embodiment A, wherein the testportion comprises at least one recessed area, wherein the testcomposition is adhered in the recessed area.

Embodiment C is the monitoring device of any one of the precedingEmbodiments, wherein the test composition further comprises a polymericbinder.

Embodiment D is the monitoring device of any one of the precedingEmbodiments, wherein the tracer analyte is selected from the groupconsisting of a plurality of viable microorganisms or a biomoleculeassociated therewith, an acid, a base, a nucleotide, a protein, anucleic acid, a carbohydrate, or hemoglobin.

Embodiment E is the monitoring device of Embodiment D, wherein theplurality of viable microorganisms comprises a yeast microorganism.

Embodiment F is the monitoring device of Embodiment D or Embodiment E,wherein the test portion comprises a porous fibrous nonwoven matrix,wherein the plurality of viable microorganisms is disposed on or in thefibrous nonwoven matrix.

Embodiment G is the monitoring device of Embodiment F, wherein the testportion further comprises a plurality of inorganic particles dispersedin the fibrous nonwoven matrix, wherein plurality of viablemicroorganisms is releasably adhered to two or more of the plurality ofinorganic particles.

Embodiment H is the monitoring device of any one of the precedingEmbodiments, further comprising a frangible seal, wherein a receivingchamber is disposed on a first side of the frangible seal proximate theopening and a cuvette chamber is disposed on a second side of thefrangible seal distal the opening.

Embodiment I is the monitoring device of Embodiment H, wherein the testelement is configured to disrupt the frangible seal.

Embodiment J is the monitoring device of any one of the precedingEmbodiments, wherein the container includes a first solvent disposedtherein.

Embodiment K is the monitoring device of Embodiment J, wherein thetracer analyte is soluble in the solvent.

Embodiment L is the monitoring device of Embodiment J or Embodiment K,wherein the test composition is dispersible in the solvent.

Embodiment M is the monitoring device of any one of the Embodiments Jthrough L as dependent on Embodiment H, wherein the solvent is disposedin the cuvette chamber.

Embodiment N is the monitoring device of Embodiment M, wherein thedetection reagent is disposed in the container.

Embodiment O is the monitoring device of Embodiment N, wherein thedetection reagent is disposed in the receiving chamber or the cuvettechamber.

Embodiment P is the monitoring device of any one of the precedingEmbodiments, wherein the test element further comprises a reservoir witha second solvent disposed therein, a hollow stem, and a liquid flowregulator capable of placing the reservoir in fluid communication withthe hollow stem.

Embodiment Q is the monitoring device of Embodiment P, wherein thetracer analyte is soluble in the second solvent.

Embodiment R is the monitoring device of Embodiment P or Embodiment Q,wherein the test composition is dispersible in the second solvent.

Embodiment S is the monitoring device of any one of Embodiments Pthrough R, wherein the detection reagent is disposed in the reservoir.

Embodiment T is the monitoring device of any one of the precedingEmbodiments, wherein the container comprises a cuvette portionconfigured to be operationally coupled with the analytical instrument.

Embodiment U is the monitoring device of any one of Embodiments Athrough T, further comprising a secural element.

Embodiment V is a method of assessing the efficacy of a washing process,comprising:

exposing the test portion of a monitoring device according to any one ofEmbodiments A through U to the washing process;

after exposing the test portion to the washing process, contacting thetest portion with the detection reagent in the container; and

using the analytical instrument to detect a presence or an absence ofthe detectable product;

wherein the presence of the detectable product indicates a presence oftracer analyte on the test portion after exposing the test portion tothe washing process.

Embodiment W is the method of Embodiment V, wherein exposing the testportion to the washing process comprises placing the test portion intoan automated washer and performing at least a portion of an automatedwash cycle while the test portion is disposed in the automated washer.

Embodiment X is the method of Embodiment W, wherein the automated washercomprises an automated washer-disinfector.

Embodiment Y is the method of any one of Embodiments V through X,wherein using the analytical instrument to detect a presence or anabsence of the detectable product comprises using the analyticalinstrument to measure a quantity of the tracer analyte.

Embodiment Z is method of any one of Embodiments W through Y, whereinexposing the test portion of a monitoring device comprises exposing thetest portion of a plurality of monitoring devices, wherein the methodfurther comprises positioning a first monitoring device at a firstpredefined location in the automated washer and positioning a secondmonitoring device at a second predefined location in the automatedwasher.

Embodiment AA is the method of Embodiment Z, as dependent uponEmbodiment Y, further comprising the step of comparing a measuredquantity of tracer analyte associated with the first monitoring deviceto a measured quantity of tracer analyte associated with the secondmonitoring device.

Embodiment BB is the method of any one of Embodiments Y through AA,further comprising comparing the measured quantity of the tracer analyteto a predefined standard.

Embodiment CC is the method of any one of Embodiments Y through BB,further comprising using the at least one measured quantity to define anaction limit.

Embodiment DD is the method of any one of Embodiments Y through BB,further comprising comparing the measured quantity to a predefinedaction limit.

Embodiment EE is the method of any one of Embodiments V through DD,further comprising associating a first datum related to a quantity oftracer analyte detected in a sample with a second datum related to otherinformation related to the sample and electronically storing theassociated first and second data.

Embodiment FF is the method of Embodiment EE, wherein the second datumcomprises information selected from the group consisting of a date, atime, a washing apparatus, an operator, an instrument to be washed, anda combination of two or more of any of the foregoing test data.

Embodiment GG is the method of any one of Embodiments V through FF,further comprising the step of placing the test portion of themonitoring device in a receiver configured to restrict fluidicaccessibility to the test portion.

Embodiment HH is the method of Embodiment GG, wherein placing the testportion of the monitoring device in a receiver configured to restrictfluidic accessibility to the test portion comprises placing the testportion into an interior space of a lumened object.

Embodiment II is the method of any one of Embodiments V through HHfurther comprising, before exposing the test portion of the monitoringdevice to the washing process, positioning at least a part of the testportion of the monitoring device in a process challenge device.

Embodiment JJ is a method of processing an object to be decontaminated,comprising:

processing as a single batch in a decontamination process:

-   -   an object having an unknown amount of biological soil disposed        thereon and/or therein;    -   a monitoring device comprising a predetermined quantity of        tracer analyte according to any one of Embodiments A-U;

after exposing the test portion to the decontamination process,contacting the test portion with the detection reagent in the container;and

using the analytical instrument to detect a presence or an absence ofthe detectable product;

wherein the presence of the detectable product indicates a presence ofthe tracer analyte on the test portion after exposing the test portionto the washing process.

Embodiment KK is the method of Embodiment JJ, wherein processing theobject and the monitoring device in a decontamination process comprisesprocessing the object and the monitoring device in an automated washeror an automated washer-disinfector.

Embodiment LL is the method of Embodiment JJ or KK, wherein processingas a single batch in a decontamination process the object and themonitoring device comprises processing as a single batch in adecontamination process the object and a plurality of the monitoringdevices.

Embodiment MM is the method of Embodiment LL, wherein processing aplurality of monitoring devices comprises processing a first monitoringdevice at a first location and processing a second monitoring device ata second location that is spaced apart from the first location.

Embodiment NN is the method of Embodiment MM as dependent on KK, whereinprocessing a plurality of monitoring devices comprises processing theplurality of monitoring devices in an automated washer or an automatedwasher-disinfector.

Embodiment OO is the method of any one of Embodiments JJ through NN,wherein detecting a presence or an absence of the detectable productfurther comprises calculating a quantity of tracer analyte remaining onthe test portion after exposing the test portion to the decontaminationprocess.

Embodiment PP is the method of Embodiment OO, further comprisingcomparing the quantity of tracer analyte remaining on the test portionof at least one monitoring device after exposing the test portion to thedecontamination process to a predetermined quantity of tracer analyteassociated with an action limit.

Embodiment QQ is the method of Embodiment PP, further comprisingreleasing the object for use in a first subsequent process if thequantity of tracer analyte remaining on the test portion of the at leastone monitoring device after exposing the test portion to thedecontamination process is less than or equal to the predeterminedquantity.

Embodiment RR is the method of Embodiment PP, further comprisingreleasing the object for use in a second subsequent process if thequantity of tracer analyte remaining on the test portion of the at leastone monitoring device after exposing the test portion to thedecontamination process is greater than or equal to the predeterminedquantity.

Embodiment SS is the method of any one of Embodiments JJ through RR,wherein calculating a quantity of tracer analyte remaining on the testportion comprises quantifying ATP, a blood component, or a protein.

Embodiment TT is a kit, comprising:

a container comprising a first end with an opening and a second endopposite the first end, wherein the container includes a detectionreagent disposed therein; and

a test element comprising a test portion to which a test composition isreleasably adhered, wherein the test composition comprises apredetermined quantity of tracer analyte;

wherein the container is configured to receive the test portion andconfigured to be operationally coupled to an analytical instrument;

wherein the tracer analyte and the detection reagent each are capable ofparticipating in one or more chemical reaction that results in theformation of a detectable product.

Embodiment UU is kit of Embodiment TT, wherein the container furthercomprises a solvent disposed therein.

Embodiment VV is the kit of Embodiment TT or Embodiment UU, wherein thetest element further comprises a reservoir with a solvent disposedtherein, wherein the test element comprises a hollow stem and thereservoir comprises a liquid flow regulator capable of placing thereservoir in fluid communication with the hollow stem.

Embodiment WW is the kit of any one of Embodiment TT through VV, furthercomprising a means to secure a test element.

Embodiment XX is the kit of any one of Embodiments TT through WW,further comprising an article comprising a receiver configured torestrict fluidic accessibility to the test portion.

Embodiment YY is the kit of Embodiment XX, wherein the article is alumened object.

Embodiment ZZ is the kit of any one of Embodiments TT through YY,wherein the tracer analyte is selected from the group consisting of aplurality of viable microorganisms, an acid, a base, a nucleotide, aprotein, a nucleic acid, a carbohydrate, or hemoglobin.

Embodiment AAA is the kit of Embodiment ZZ, wherein the test portioncomprises a fibrous nonwoven matrix, wherein the plurality of viablemicroorganisms is disposed on or in the fibrous nonwoven matrix.

Embodiment BBB is the kit of Embodiment AAA, wherein the test portionfurther comprises a plurality of inorganic particles dispersed in thefibrous nonwoven matrix, wherein the plurality of viable microorganismsis releasably adhered to two or more of the plurality of inorganicparticles.

Embodiment CCC is a system for testing the efficacy of a washingprocess, comprising:

a monitoring device comprising a container and a test element thatincludes a test portion to which a test composition comprising apredetermined quantity of a tracer analyte is releasably adheredaccording to any one of Embodiments A through U; and

an analytical instrument capable of detecting the detectable product;

wherein the container is configured to receive the test portion andconfigured to be operationally coupled to the analytical instrument.

Embodiment DDD is the system of Embodiment CCC, further comprising acomputer capable of receiving data from the analytical instrument and amemory capable of storing the received data.

Embodiment EEE is the system of Embodiment CCC or Embodiment DDD,further comprising a process challenge device capable of receiving atleast a part of the test portion of the test element.

Embodiment FFF is an article comprising a homogeneous dried compositionremovably adhered thereto, wherein the composition comprises apredetermined amount of adenosine-5′-triphosphate and a dye in an amountsufficient to be optically detectable.

Embodiment GGG is the article of Embodiment FFF, wherein the compositionfurther comprises a polymeric binder.

Embodiment HHH is the article of Embodiment GGG, wherein the polymericbinder comprises a water-dispersible polymeric binder.

Embodiment III is the article of Embodiment GGG or Embodiment HHH,wherein the polymeric binder comprises polyvinyl alcohol, polyethyleneglycol, or mixtures thereof.

Embodiment JJJ is a homogeneous, dried artificial test soil consistingessentially of adenosine-5′-triphosphate, or a salt thereof, and apolymeric binder.

Embodiment KKK is the test soil of Embodiment JJJ, wherein the polymericbinder comprises polyvinyl alcohol, polyethylene glycol, or mixturesthereof.

Embodiment LLL is the test soil of Embodiment JJJ or Embodiment KKK,wherein the polymeric binder comprises a water-dispersible polymericbinder.

Objects and advantages of this invention are further illustrated by thefollowing examples, but the particular materials and amounts thereofrecited in these examples, as well as other conditions and details,should not be construed to unduly limit this invention.

EXAMPLES

Unless stated otherwise, all reagents used in the following exampleswere reagent-grade. 3M CLEAN-TRACE Water Test (Total ATP) test units, 3MCLEAN-TRACE Water Test (Free ATP) test units, and the 3M CLEAN-TRACE NGLuminometer were obtained from 3M Company (St. Paul, Minn.).

Preparative Example 1—Preparation of Tracer Analyte-Coated Test Elements

A dilute suspension of porcine red blood cells (Solution I) was preparedby mixing 50 milliliters of porcine blood in 0.4% (w/v) sodium citrate(obtained from Cocalico Biological, Inc.; Reamstown, Pa.) with 50milliliters of 0.9% (w/v) sodium chloride. A solution (Solution II) of250 mM calcium chloride was prepared by dissolving 1.387 g of CaCl₂) indistilled water. A coating solution (Solution III) was prepared bymixing 20 milliliters of Solution I with 0.2 milliliters of Solution II.Solution III was used to coat sample acquisition devices less than 15minutes after it was prepared because the addition of the calciumchloride caused the blood to clot.

The sample-acquiring tips of five 3M CLEAN-TRACE Water Test (Total ATP)sample acquisition devices (Set A) were dipped into Solution III andimmediately withdrawn from the coating solution. In addition, thesample-acquiring tips of five 3M CLEAN-TRACE Water Test (Free ATP)sample acquisition devices (Set B) were dipped into Solution III andimmediately withdrawn from the coating solution. All coated sampleacquisition devices (hereinafter, “test elements”) were inverted and thehandles were placed into a microcentrifuge rack. The test elements wereallowed to dry in a biosafety hood at ambient temperature and humidityfor 2.5 hours. After drying, the test elements were returned to theirrespective ATP test units.

Preparative Example 2—Preparation of Corona-Treated, TracerAnalyte-Coated Test Elements

In order to increase the hydrophilicity of the coated portion of thetest elements, representative sets of sample acquisition devices weresubjected to corona plasma treatment before dipping each of them into ablood-containing solution and drying them as described in PreparativeExample 1.

3M CLEAN-TRACE Water Test (Total ATP) sample acquisition devices (Set C)were surface-treated using a corona plasma process. The devices wereheld by the handle and the tips were positioned about 1.4 mm away from ahand-held corona plasma source (model BD-20AC corona plasma sourceobtained from Electro-Technic Products, Inc.; Chicago, Ill.). Thedevices were slowly rotated about their respective longitudinal axis forabout 1 minute, thereby exposing the entire circumference of the tip tothe corona plasma field. The devices were subsequently dipped intoSolution III and dried as described in Preparative Example 1.

Preparative Example 3—Preparation of Diamond-Like Glass-Coated, TracerAnalyte-Coated Test Elements

In order to increase the hydrophilicity of the coated portion of thetest elements, representative sets of sample acquisition devices werecoated with diamond-like glass before dipping each of them into ablood-containing solution and drying them as described in PreparativeExample 1.

3M CLEAN-TRACE Water Test (Total ATP) sample acquisition devices (Set D)were coated with a diamond-like glass coating according to the followingprocess. The diamond-like glass film was deposited in a commercial batchreactor (Plasmatherm Model 3032), which was configured for reactive ionetching (RIE) with a 69 cm lower powered electrode and central gaspumping. The chamber was pumped by a roots blower (Edwards Model EH1200)backed by a dry mechanical pump (Edwards Model iQDP80). RF power wasdelivered by a 5 kW, 13.56 Mhz solid-state generator (RFPP Model RF30Sthrough an impedance matching network. The system had a nominal basepressure of 0.667 Pascal. The flow rates of the gases were controlled bymass flow controllers (MKS Instruments, Inc.). Substrates for depositionwere placed on the lower powered electrode. The samples were then plasmatreated in the following manner. The samples were placed on the poweredelectrode of the batch plasma apparatus. The plasma treatment was donein a series of treatment steps. First, the tips were treated with oxygenplasma by flowing oxygen gas at a flow rate of 500 standard cm3/min andplasma power of 300 watts for 20 seconds. After the oxygen plasmatreatment, a diamond-like glass film was deposited by flowingtetramethylsilane gas at a flow rate of 150 standard cm3/min, and plasmapower of 300 watts for 20 seconds. The diamond-like glass film wasfurther surface modified by treating in oxygen plasma at a flow rate of500 cm3/min and plasma power of 300 watts for 60 seconds. After theplasma deposition was completed, the chamber was vented to atmosphereand the samples were removed.

The devices were subsequently dipped into Solution III and dried asdescribed in Preparative Example 1.

Preparative Example 4—Preparation of Surface-Treated, TracerAnalyte-Coated Test Elements

3M CLEAN-TRACE Water Test (Total ATP) sample acquisition devices (Set G)were surface-treated using a corona plasma process similar to thatdescribed in Preparative Example 2 except the sample acquisition deviceswere rotated in the corona plasma for an additional 30 seconds. Thedevices were subsequently dipped into Solution III and dried asdescribed in Preparative Example 1.

Preparative Examples 5-7 and 5a-7a—Preparation of Test CompositionsComprising Adenosine-5′-Triphosphate and a Polymeric Binder

CELVOL-brand polyvinyl alcohol polymers were obtained from SekisuiSpecialty Chemicals (Secaucus, N.J.). A stock solution (1 mg/mL) of ATPwas prepared in sterile deionized water. The stock solution was seriallydiluted in sterile deionized water to produce one working solution (“A”)containing 1 microgram/mL ATP and another working solution (“B”)containing 0.7 micrograms/mL ATP, respectively. For Preparative Examples5-7, aliquots of working solution “A” were added to individual mixingjars. For Preparative Examples 5a-7a, aliquots of working solution “B”were added to individual mixing jars. A solution of FD&C Red Dye #40 wasprepared by dissolving 160 mg of F&DC Red Dye #40 into 40 mL of sterilewater. The 4 mg/mL Red Dye #40 solution was added to each of the jarswith the ATP working solution and the jars were placed in a water bathat 80° C. The appropriate amount of a designated polymeric binder(reported in each particular Example, below) was added to each jar at arate of about 1.0 gram/minute with stirring to obtain the reportedconcentration of binder. Each mixture was stirred for about one hour toallow the polymeric binder to fully dissolve. The final concentration ofATP for each Preparative Example 5-7 was 1 microgram/mL. The finalconcentration of Red Dye #40 was 0.13 mg/mL, which was added primarilyfor visibility when coating the device tips.

TABLE 1 Composition of Artificial Test Soils comprising ATP and apolymeric binder. Preparative Example Polymeric Binder ATP Concentration5 Polyvinyl alcohol (CELVOL 425) 1.0 microgram/mL 5a Polyvinyl alcohol(CELVOL 425) 0.7 microgram/mL 6 Polyvinyl alcohol (CELVOL 443) 1.0microgram/mL 6a Polyvinyl alcohol (CELVOL 443) 0.7 microgram/mL 7Polyethylene glycol (8000) 1.0 microgram/mL 7a Polyethylene glycol(8000) 0.7 microgram/mL

Preparative Example 8—Preparation of Artificial Test Soils (ATSs)Comprising Adenosine-5′-Triphosphate

A commercially available dehydrated artificial test soil with theproduct name WASHER DISINFECTOR SOIL TEST (Order Code 2304), used forPreparative Example 8 was obtained from Albert Browne International Ltd(Leicester, UK). The artificial test soil was rehydrated using a sterilewater solution containing 1 microgram/mL ATP that was made as describedin Preparative Examples 5-7.

Example 1—Measurement of ATP from Blood-Coated Test Elements

The amount of ATP from the blood coated on each test element madeaccording to Preparative Example 1 was measured in a bioluminescent(i.e., luciferin/luciferase reaction) assay using a 3M CLEAN-TRACE NGLuminometer. At least portions of the coating were observed to crack andseparate from the test elements after drying. Table 2 shows the ATPmeasurement results from each set.

TABLE 2 Amount of ATP detected from blood-coated test elements. Allresults are reported in Relative Light Units (RLU). Test Element Set A(Total ATP) Set B (Free ATP) (n = 4) (n = 5) Average 1,104,693 1,097,112STDEV +/−29,179 +/−14,710

Example 2—Measurement of ATP from Surface-Treated, Blood-Coated TestElements

3M CLEAN-TRACE Water Test (Total ATP) sample acquisition devices (Set D)were surface-treated using a diamond-like glass (DLG) coating process.The devices were coated with DLG using the method described above. Afterthe devices were coated with the DLG, they were dipped in Solution IIIand dried as described in Preparative Example 1.

3M CLEAN-TRACE Water Test (Total ATP) sample acquisition devices (Set E)did not receive any surface treatment. They were dipped in Solution IIIand dried as described in Preparative Example 1.

A 3M CLEAN-TRACE Water Test (Total ATP) sample acquisition device (SetF) did not receive any surface treatment. The device was dipped inSolution I (Example 1) and dried according to the process described inPreparative Example 1.

The blood-coated devices (i.e., test elements) were returned to theirrespective ATP test units and the amount of ATP from the blood coated oneach test element was measured in a bioluminescent (i.e.,luciferin/luciferase reaction) assay using a 3M CLEAN-TRACE NGLuminometer. Table 3 shows the ATP measurement results from each set. Itwas noted that, although some flaking of the dried coating occurred ontest elements from Set C and Set D, it was significantly less than setE. indicating the coating was retained better by the surface-modifiedtest elements than by the test elements that were not surface-treated.

TABLE 3 Amount of ATP detected from blood-coated test elements. Allresults are reported in Relative Light Units (RLU). The number shown inparentheses is the number of test elements that was tested for eachparticular set. Test Element Set C (n = 3) Set D (n = 4) Plasma CoronaDLG Coated Set E (n = 2) Set F (n = 1) Treated Dipped in Dipped inUntreated Untreated Soln. III Soln. III Soln. III Soln. I Average1,150,369 1,108,209 1,145,357 1,153,984 STDEV +/−10965 +/−71931 +/−18705(n = 1)

Example 3—the Use of Test Elements to Monitor the Efficacy of aDefective Washing Process

Test elements made according to Preparative Example 3 were placed intostainless steel wire mesh baskets (approximate size 50 cm long×20 cmwide×10 cm tall) that are used to hold articles to be washed in aGETINGE 46-4 model-washer disinfector (Getinge USA, Inc., Rochester,N.Y.). All test elements of Sets C and D were securely fastened to theinside corners of the baskets using hose clamps and zip ties. The testelements of Set E were placed at one end inside each basket. The basketswere then placed into the GETINGE 46-4 instrument washer. Two basketswere placed in each of the three levels available in the GETINGE 46-4instrument washer. A “defective” washer-disinfector cycle was run. Thedefective cycle included a shortened hot-water (60° C.) wash, did notinclude use of a detergent, and did not include a very-hot (90° C.)final rinse. The parameters for this defective wash cycle (“IncompleteCycle I”) are listed in Table 4. The conditions of the wash cycle werenot sufficient to comply with most existing standards for washingmedical instruments in a hospital. After completion of the wash cycleand prior to the measuring the blood (ATP) retained on each testelement, residual water from the wash cycle was removed from each tip oftest elements in a first group (Group A), by gently shaking the testelement 2 or 3 times. The other test elements (Group B) were not shakenprior to ATP measurement. The washed test elements were returned totheir respective ATP test units and the amount of ATP from the bloodcoated on each test element was measured in a bioluminescent (i.e.,luciferin/luciferase reaction) assay using a 3M CLEAN-TRACE NGLuminometer. Table 5 shows the ATP measurement results from each set.

TABLE 4 Washing, rinsing, and drying parameters for Incomplete Cycle I.Length Process Step 1.0 min. Cold-water pre-rinse 3.0 min. Hot-water(60° C.) wash 1.0 min. Hot-water (60° C.) first rinse 1.0 min. Hot-water(60° C.) second rinse 10.0 min.  Dry

TABLE 5 Amount of ATP detected from blood-coated test elements. Allresults are reported in Relative Light Units (RLU). Negative Controldevices that were not surface-modified and were not dipped in swineblood had readings of 4 RLU to 12 RLU (data not shown in table). Set CSet D Set E Plasma Corona DLG Treated Untreated Treated Dipped DippedDipped in Soln. III in Soln. III in Soln. III Group A 570 327 122(shaken) 49 255 Group B 99 490 67 (not shaken) 808 436 Average 382 37795 STDEV +/−369 +/−106 39

Example 4—the Use of Test Elements to Monitor the Efficacy of aNondefective Washing Process

Test elements made according to Preparative Example 4 (Set G) and Set E(described above) were placed into stainless steel wire mesh basketsthat are used to hold articles to be washed in a GETINGE 46-4 modelwasher disinfector (Getinge USA, Inc., Rochester, N.Y.). All testelements of Set G were securely fastened to the inside corners of thebaskets using hose clamps and zip ties. The test elements of Set E werelikewise secured at one end inside each basket. Additionally a set ofnegative controls were placed loosely on the bottom of each basket.These negative control devices were not surface-modified and were notdipped in swine blood. The baskets were then placed into the GETINGE46-4 instrument washer. A “nondefective” washer-disinfector cycle wasrun. The nondefective cycle included a longer hot-water (60° C.) washthan the “defective cycle, it include use of a multi-enzyme detergent(BMEC 70508-A detergent available from 3M Company, St. Paul, Minn.), andit included a very-hot (90° C.) final rinse. The parameters for thisnondefective wash cycle (“Complete Cycle II”) are listed in Table 6. Theconditions of the wash cycle were sufficient to comply with mostexisting standards for washing medical instruments in a hospital. Aftercompletion of the wash cycle and prior to the measuring the blood (ATP)retained on each test element, residual water from the wash cycle wasremoved from each tip of test elements by gently shaking the testelement 2 or 3 times. The washed test elements were returned to theirrespective ATP test units and the amount of ATP from the blood coated oneach test element was measured in a bioluminescent (i.e.,luciferin/luciferase reaction) assay using a 3M CLEAN-TRACE NGLuminometer. Table 7 shows the ATP measurement results from each set.

TABLE 6 Washing, rinsing, and drying parameters for Complete Cycle II.Length Process Step 1.0 min. Cold-water pre-rinse 5.0 min. Hot-water(60° C.) wash with detergent 1.0 min. Hot-water (60° C.) first rinse 1.0min. Hot-water (60° C.) second rinse 1.0 min. Very-hot (90° C.) finalrinse 10.0 min.  Dry

TABLE 7 Amount of ATP detected from blood-coated test elements. Allresults are reported in Relative Light Units (RLU). The number shown inparentheses is the number of test elements that was tested for eachparticular set. Negative Control devices were not surface-modified andwere not dipped in swine blood. Set G Set E Plasma Corona UntreatedNegative Control Treated Dipped in Dipped in Untreated Soln. III Soln.III No Soil (n = 8) (n = 3) (n = 4) Average 16 7 8 STDEV +/−6 +/−1 +/−3

The corona-treated test devices from Set G that were exposed to theComplete Cycle II shown in Table 7, all showed lower levels of ATP thanthe corresponding test devices that were exposed to the Incomplete CycleI (see Table 5). In addition, the ATP levels measured from blood-coated,corona-treated test devices exposed to Complete Cycle II were onlyslightly higher than the negative control test devices, which were notcoated with swine blood.

Example 5—Measurement of ATP from Surface-Treated Test Elements withCELVOL Polymeric Binder

CLEAN-TRACE Water Test (Total ATP) test units were plasma corona treatedas described above for 90 seconds. The devices were subsequently dippedinto Preparative Example 5a (CELVOL 425), to create Set H. Other plasmacorona treated devices were subsequently dipped into Preparative Example6a (CELVOL 443), to create Set I. After dipping, test devices of Set Hand Set I were rotated slowly on top of a small piece of PARAFILM toremove excess material of the ATP spiked binder coating from the tip ofthe test device. The test devices were dried for 1 hour at 60 degreesC., then shaken by hand for 45 seconds, and then to returned to theirrespective ATP test units and the amount of ATP on each test element wasmeasured in a bioluminescent (i.e., luciferin/luciferase reaction) assayusing a 3M CLEAN-TRACE NG Luminometer. Table 8 shows the ATP measurementresults from each set. Additionally, for comparison, 3 test devices forSet H were also assessed for ATP without drying, in a “wet” condition.

TABLE 8 Amount of ATP detected from CELVOL Polymeric Binder coated testelements. All results are reported in Relative Light Units (RLU). Thenumber shown in parentheses is the number of test elements that wastested for each particular set. Set H Set H Set I 13.0 wt. % 13.0 wt. %14.9 wt. % CELVOL 425 CELVOL 425 CELVOL 443 Wet (not dried) Dried 1 hourDried 1 hour (n = 3) (n = 3) (n = 3) Average 611,736 412,296 402,364STDEV +/−49,332 +/−30,452 +/−60,943

Example 6—Measurement of ATP from Surface-Treated Test Elements with ATPSpiked Coatings after Wash Disinfection Cycle

CLEAN-TRACE Water Test (Total ATP) test units were plasma corona treatedas describe above for 90 seconds. The devices were subsequently dippedinto the following Preparative Examples containing ATP: PreparativeExample 6a (CELVOL 443), to create Set J, or dipped in PreparativeExample 7a (PEG) to create Set K or dipped in Preparative Example 8(commercially available artificial test soil), to create Set L. Theconcentration of the binder portion of Preparative Examples 6 and 7 aredescribed in Table 8, below. After dipping, test devices of Sets J, Kand L were brushed with a small commercially available acid brush in adirection parallel to the major axis of the test device, which is adirection orthogonal to the recessed grooves of the test device. Thisbrushing physically removed the excess material of the ATP spikedcoating from the tip of the test device, leaving the majority of thecoated material remaining within the recessed grooves of the test devicetip. The test devices were dried for 1 hour at 60° C., and then shakenby hand for 45 seconds. Each of Set J, K and L were divided into 3sub-groups. The 3 sub-groups were subjected to one of (1) “No Wash” or(2) the defective wash cycle “Incomplete Cycle I” described in Table 4,or (3) the nondefective wash cycle, “Complete Cycle II” described inTable 6. Each sub-group under each wash condition, including a set ofuncoated test devices as controls, consisted of 5 individual testdevices (n=5). The monitoring devices that were to be subjected to washcycles were placed loosely in the corner of the stainless steel wiremesh baskets that are used to hold articles to be washed in a GETINGE46-4 model washer disinfector, used in Example 4. Afterwards themonitoring devices were then to returned to their respective ATP testunits and the amount of ATP on each test element was measured in abioluminescent (i.e., luciferin/luciferase reaction) assay using a 3MCLEAN-TRACE NG Luminometer. Table 9 shows the ATP measurement resultsfrom each J, K and L set and sub-group.

TABLE 9 Amount of ATP detected from coated test elements. All resultsare reported in Relative Light Units (RLU). Negative Control deviceswere not surface-modified and were not coated. Set J Negative 9.1 wt. %Set K Controls CELVOL 50.0 wt. % Set L Uncoated 433 PEG ATS Test UnitsConditions (n = 5) (n = 5) (n = 5) (n = 5) No Wash Average 560,327557,192 16,852 182 No Wash STDEV +/−10,104 +/−48,598 +/−2,159 +/−294Cycle I Average 3,088 949 1,515 52 Cycle I STDEV +/−3,589 +/−1,715+/−3,235 +/−8 Cycle II Average 527 34 107 24 Cycle II STDEV +/−1,110+/−18 +/−191 +/−28

Example 7—Measurement of ATP from Surface-Treated Test Elements with ATPSpiked Coatings after Extended Storage and Wash Disinfection Cycles

Monitoring devices were prepared as in Example 6, Set J, dipped intoPreparative Example 6a (CELVOL 443, and brushing with the acid brush, tocreate Set M. However, additionally for Set M, the test devices weredipped and brushed a total of four times (4 coatings). The intermittentdrying time was 40 minutes in a 60° C. oven, followed by a final drytime of 1 hour at 60° C. The concentration of the CELVOL 443 portion ofPreparative Example 6a used for Set M was 9.2 wt. %. Set M was dividedinto sub-groups that were stored under ambient conditions for variouslengths of time (0, 7, 14 days) to evaluate the stability over time ofthe coated monitoring devices prior to use. Negative Controls (untreatedand uncoated test devices) were stored under like conditions over thesame time periods. Also like Example 6, the sub-groups of Set M werealso subjected to one of (1) “No Wash” or (2) the defective wash cycle“Incomplete Cycle I” described in Table 4, or (3) the nondefective washcycle, “Complete Cycle II” described in Table 6. The test devices thatwere to be subjected to wash cycles were placed loosely in the corner ofthe stainless steel wire mesh baskets that are used to hold articles tobe washed in a GETINGE 46-4 model washer disinfector, used in Example 4.Afterwards the devices were then to returned to their respective ATPtest units and the amount of ATP on each test element was measured in abioluminescent (i.e., luciferin/luciferase reaction) assay using a 3MCLEAN-TRACE NG Luminometer. Table 10 shows the ATP measurement resultsfrom Set M.

TABLE 10 Amount of ATP detected from 4X coated test elements afterstorage. All results are reported in Relative Light Units (RLU).Negative Control devices were not surface-modified and were not coated.Set M Set M Set M Day 0 Day 7 Day 14 Conditions (n = 4) (n = 4) (n = 5)No Wash Average 782,458 750,544 750,156 No Wash STDEV +/−16,528+/−27,419 +/−15,863 No Wash Neg. Control Average 11 14 70 No Wash Neg.Control STDEV +/−2 +/−1 +/−27 Cycle I Average 3,461 707 1,299 Cycle ISTDEV +/−5,002 +/−166 +/−1,430 Cycle I Neg. Control Average 164 100 78Cycle I Neg. Control STDEV +/−189 +/−31 +/−32 Cycle II Average 45 60 45Cycle II STDEV +/−5 +/−20 +/−12 Cycle II Neg. Control Average 15 — —Cycle II Neg. Control STDEV +/−8 — —

Example 8—Measurement of ATP from Surface-Treated Test Elements with ATPSpiked Coatings after Hospital Wash Disinfection Cycles

Monitoring devices were prepared as in Example 7, four coatings ofPreparative Example 6a (CELVOL 443), and brushing with the acid brush,to create Set N. The concentration of the CELVOL 443 portion ofPreparative Example 6a used for Set N was 9.7 wt. %. Set N was dividedmultiple sub-groups that were used to assess the wash disinfectingcycles of hospital grade commercial machines. Four different machines,all of which were GETINGE DISINFECTION DECOMATT 8666WASHER-DISINFECTORS, were used to process 9 loads of instruments usingtwo different wash disinfection cycles. The “P1” cycle represented a“normal” load of surgical instruments and a regular wash cycle. The “P2”cycle represented a heavy duty wash cycle and contained surgicalinstruments that were typically more soiled than those in the “normal”load. The monitoring devices were placed in baskets with the surgicalinstruments and the baskets were placed into the washer disinfectionmachines in one of 4 locations: T=top rack, 2=second rack down, 3=thirdrack down, and B=bottom rack. Additionally, Negative Control testdevices (untreated and uncoated) were run through the same cycles as SetN. After the wash disinfection cycles, the test devices were removedfrom the baskets and were then returned to their respective ATP testunits and the amount of ATP on each test element was measured in abioluminescent (i.e., luciferin/luciferase reaction) assay using a 3MCLEAN-TRACE NG Luminometer. Table 11 shows the ATP measurement resultsfrom Set N coated monitoring devices. Table 12 shows the ATP measurementresults from the Negative Control test devices, processed with Set N.

TABLE 11 Set N - Amount of ATP detected from 4X coated test elementsafter Wash Disinfection Cycle in Hospital size Equipment. All resultsare reported in Average Relative Light Units (RLU). Ma- Load chine CycleT 2 3 B 1 4 P1 731 2,404 3,786 474 2 2 P1 28,456 2,764 4,342 2,200 3 1P2 3,618 3,393 3,670 2,393 4 4 P2 338 2,816 5,494 2,776 5 2 P2 2,8223,287 1,768 306 6 3 P2 11,484 8,196 6,609 6,053 7 1 P2 2,591 2,723 2,5122,080 8 3 P2 3,032 3,247 4,775 4,154 9 1 P1 8,160 1,441 1,330 1,594Average — — 6,804 3,363 3,698 2,448 STDEV — — +/−8,869 +/−1,906 +/−1,771+/−1,783

TABLE 12 Amount of ATP detected from Negative Control test devices afterWash Disinfection Cycle in Hospital size Equipment. All results arereported in Average Relative Light Units (RLU). Load Machine Cycle T 2 3B 1 4 P1 17 17 30 73 2 2 P1 50 75 54 169 3 1 P2 45 57 13 15 4 4 P2 23 39132 18 5 2 P2 43 305 58 65 6 3 P2 83 63 142 66 7 1 P2 37 28 87 119 8 3P2 63 130 124 116 9 1 P1 46 20 45 36 Average — — 45 82 76 75 STDEV — —+/−20 +/−91 +/−41 +/−51

Example 9—Measurement of ATP from Surface-Treated Test Elements with ATPSpiked Coatings after Wash Disinfection Cycle

Three different test devices were coated with the Preparative Example 6a(CELVOL 443) according to the following procedures. Set 0 test deviceswere created by taking 3M CLEAN-TRACE SURFACE PROTEIN HIGH SENSITIVITYswab Cat # MPRO50, available from 3M Company of St. Paul, Minn., andpipetting onto it 0.2 mL of 0.7 micrograms/mL of ATP in a 9.7 wt. %CELVOL binder, otherwise prepared according to Preparative Example 6 anddried for 1 hour at 60° C. Set P test devices were created by taking 3MCLEAN-TRACE SURFACE ATP swab Cat # UXL100 (a rayon material swab),available from 3M Company, and pipetting onto it 0.2 mL of 0.7micrograms/mL of ATP in a 9.7 wt. % CELVOL binder, otherwise preparedaccording to Preparative Example 6a and dried for 1 hour at 60° C. Set Qtest devices were created by taking 3M CLEAN-TRACE SURFACE ATP swab Cat# UXL100, available from 3M Company, removing the swab tip (leaving onlythe “stick” portion of the device) and dip coating four timesapproximately 1.3 cm of the end of the stick with Preparative Example 6asolution containing 0.7 micrograms/mL of ATP in a 9.7 wt. % CELVOLbinder and dried for 1 hour at 60° C. Each of Set O, P and Q weredivided into 3 sub-groups. The 3 sub-groups were subjected to one of (1)“No Wash” or (2) the defective wash cycle “Incomplete Cycle I” describedin Table 4, or (3) the nondefective wash cycle, “Complete Cycle II”described in Table 6. The test devices that were to be subjected to washcycles were placed loosely in the corner of the stainless steel wiremesh baskets and processed the in a GETINGE 46-4 model washerdisinfector described in Example 4. Afterwards the test devices werethen to returned to their respective ATP test units and the amount ofATP on each test element was measured in a bioluminescent (i.e.,luciferin/luciferase reaction) assay using a 3M CLEAN-TRACE NGLuminometer. Table 13 shows the ATP measurement results of Sets O, P andQ.

TABLE 13 Amount of ATP detected from coated test elements of variouscompositions. All results are reported in Relative Light Units (RLU).Set O Set P Set Q 9.7 wt. % 9.7 wt. % 9.7 wt. % CELVOL 433 CELVOL 433CELVOL 433 Conditions MPRO50 swabs UXL100 swabs Plain “sticks” No Wash450,666 (n = 4) 434,232 (n = 4) 82,495 (n = 10) Average No Wash 27,43318,468 12,392 STDEV Cycle I 5,137 (n = 5) 3,745 (n = 5) 19,963 (n = 8)Average Cycle I 2,261 2,929 19,796 STDEV Cycle II 433 (n = 5) 425 (n =4) 7,518 (n = 10) Average Cycle II 200 126 9,467 STDEV

Example 10. Measurement of ATP from Surface-Treated Test Elements withATP Spiked Coatings after Hospital Wash Disinfection Cycles

Monitoring devices were prepared as in Example 7, using four coatings ofPreparative Example 6a (CELVOL 443) and brushing with the acid brush tocreate Set 1. The concentration of the CELVOL 443 portion of PreparativeExample 6a used for Set 1 was 9.7 wt. %. Set 1 was divided multiplesub-groups that were used to assess the wash disinfecting cycles ofhospital grade commercial machines. Two different machines, both ofwhich were STERIS RELIANCE 444 WASHER DISINFECTOR, were used to process9 loads of instruments using two different wash disinfection cycles. The“Instrument” cycle represented a “normal” load of surgical instrumentsand a regular wash cycle. The monitoring devices were placed in basketswith the surgical instruments and the baskets were placed into thewasher disinfection machines in one of 4 locations: T=top rack, 2=secondrack down, 3=third rack down, and B=bottom rack. After the washdisinfection cycles, the test devices were removed from the baskets andwere then returned to their respective ATP test units and the amount ofATP on each monitoring device test element was measured in abioluminescent (i.e., luciferin/luciferase reaction) assay using a 3MCLEAN-TRACE NG Luminometer. Table 14 shows the ATP measurement resultsfrom Set 1 coated monitoring devices.

TABLE 14 Set 1 - Amount of ATP detected from quadruple-coated testelements after Wash Disinfection Cycle in Hospital size Equipment. Allresults are reported in Average Relative Light Units (RLU). Load MachineCycle T 2 3 B 1 1 Instrument 1,604 1,081 334 600 2 1 Instrument 1,331860 1,345 8,035 3 1 Instrument 820 302 1,254 1,244 4 2 Instrument 205208 258 273 5 2 Instrument 9,374 386 273 351 6 2 Instrument 242 201 474201 Average — — 2,263 506 656 1,784 STDEV — — ±3,529 ±372 ±504 ±3,085

TABLE 15 Set 1 - Mean and upper control limit determined for each washerdisinfector machine. All results are reported in Average Relative LightUnits (RLU). The upper control limit in this instance was calculated asthe mean plus three standard deviations. The calculated mean andstandard deviation are shown in Table 14. Table 15. Set 1 - Mean andupper control limit determined for each washer disinfector machine. Allresults are reported in Average Relative Light Units (RLU). UpperControl Limit Machine Cycle Mean (Mean + 3σ) 1 Instrument 1,047 12,706 2Instrument 268 681

A two-sample T-test of the data comparing Machine 1 to Machine 2resulted in a P-value of <0.001.

TABLE 16 Materials for Examples 11-12. CM-111 Amorphous magnesiumsilicate purchased from 3M Company, St. Paul, MN, as 3M CosmeticMicrospheres CM-111 YM plate yeast and mold detection plate, obtainedfrom 3M Company, St. Paul, MN, under the trade designation 3M PETRIFILMYEAST AND MOLD PLATE YPD agar plate agar plate prepared according tomanufacturer's instructions with 5 wt % Yeast Extract Peptone Dextroseand 1.5 wt % agar, both powders from BD (DIFCO), Sparks MD DI waterdeionized, filtered, 18 megaohm water, processed through Milli-QGradient System obtained from Millipore; Waltham, MA Fiber 1 1 denierfibrillated polyethylene fibers, obtained from Minifibers, Inc., JohnsonCity, TN, under the trade designation FYBREL600 Fiber 2 6 denier 2inches long chopped nylon fibers, obtained from Minifibers, Inc. Fiber 31 denier bicomponent ethylene vinyl acetate/polypropylene fibers,obtained from Minifibers, Inc. Fiber 4 long glass fibers, obtained fromSchuller, Inc., Denver, CO, under the trade designation MICRO-STRAND106-475 GLASS MICROFIBERS Flocculant flocculant agent obtained fromMidsouth Chemical Co., Inc., Ringgold, LA, under the trade designation9307 FLOCCULANT Latex binder 50% solids vinyl acetate emulsion, obtainedfrom Air Products Polymers, Allentown, PA, under the trade designationAIRFLEX 600BP Saccharomyces Purchased from American Type CultureCollection, Manassas, VA cerevisiae ATCC 201390 Detergents GETINGEAlkaline cleaner detergent, catalog number 61301605277 used (40 mL/4gallons); GETINGE Dual enzyme instrument detergent, catalogue number61301605269 (40 mL/4 gallons)

Example 11—Preparation of Carrier Material A

A fiber premix was prepared by mixing 67.50 grams of Fiber 1, 13.50grams of Fiber 2, 10.13 grams of Fiber 3, and 7.87 grams of Fiber 4 with4 liters of cold tap water in a 4 L blender (available from VWR, Radnor,Pa., under the trade designation WARING COMMERCIAL HEAVY DUTY BLENDER,MODEL 37BL84) at medium speed for 30 seconds. The mixture was examinedfor uniform dispersion of the fibers without nits or clumps, and blendedfurther for 15 seconds on low speed to break up clumps. The fiber premixwas added to a 10 liter stainless steel beaker and mixed with animpeller mixer (obtained from ThermoFisher Scientific, Waltham, Mass.,under the trade designation STEDFAST STIRRER MODEL SL2400) at a speedsetting of 4 for five minutes. Then 0.6 grams of latex binder wasdispersed in about 25 mL of tap water in a 50 mL beaker and added to themixture. The beaker was rinsed with about another 25 mL of tap waterthat was added to the mixture and mixed for about 2 minutes. An amountof 22.5 grams of CM-111 powder was added to the mixture and mixed forabout 1 minute. In the same manner as latex binder, 0.6 grams offlocculant was dispersed in about 25 mL of tap water and added to themixture while mixing, followed by the addition of about another 25 mL ofrinse water from the beaker. The latex binder crashed out of solutiononto the fibers and the liquid phase of the premix changed from cloudyto substantially clear.

A felt of the above material was prepared using a pad maker apparatus(obtained from Williams Apparatus, Watertown, N.Y., under the tradedesignation TAPPI). The pad maker had a box measuring about 30centimeters (12 inches) square and 30 centimeters (12 inches) high witha fine mesh screen at the bottom and a drain valve. The box was filledwith tap water up to a height of about 1 centimeter above the screen.The mixture containing CM-111 was poured into the box and the valve wasopened immediately which created a vacuum that pulled the water out ofthe box. The resulting wet-laid felt was approximately 3 millimetersthick. The wet-laid felt was transferred from the apparatus onto a 20centimeter square sheet of blotter paper (96-pound white paper, obtainedfrom Anchor Paper, St. Paul, Minn.). The felt was sandwiched between 2to 4 layers of blotter paper, to blot excess water. The pressed felt wasthen transferred onto a fresh sheet of blotter paper and placed in anoven (obtained from SPX Thermal Product Solutions, White Deer, Pa.,under the trade designation BLUE M STABIL-THERM OVEN, MODEL OV-560A2)set at 110° C. for about 2 hours to remove residual water and cure thelatex binder to form a porous matrix.

Example 12—Preparation of Carrier Material B

Example 12 was another fiber premix and felt prepared according to theprocedure described for Example 11, above, except that the fiber premixcontained 25 grams of Fiber 1, 5.06 grams of Fiber 2, 3.80 grams ofFiber 3, and 2.95 grams of Fiber 4 and the amount of CM-111 was 8.44 gto form carrier material B.

Preparation of Yeast Stock Solution

A single isolated colony from a streak culture of Saccharomycescerevisiae (S. cerevisiae) from a YPD agar plate was used to inoculate 5mL yeast extract potato dextrose broth (YPD Broth prepared 5% w/v,purchased from Becton Dickenson, Sparks, Md.) and incubated overnight at30° C.

Spiking Carrier Material A and B with Yeast

Several 6 mm diameter disks, were die-punch cut from the carriermaterial A (Example 11) and carrier material B (Example 12), describedabove. A 100 microliter volume from the overnight yeast stock solution,containing approximately 1×10⁸ CFU/mL, was added on top of each of thesample disks of Example 11 and Example 12. The resulting yeast spikeddisks of Example 11 and Example 12 contained approximately 1.8×10⁷CFUs/mL of S cerevisiae (based on colony counts on YM Plate), and werestored in a sterile petri dish at room temperature (about 25° C.)overnight.

Mounting Yeast Spiked Disks of Example 11 and Example 12

Disks of Example 11 and Example 12, all spiked with yeast, were mountedinto one of two different types of monitoring devices. Monitoring device#1 was created by utilizing the plastic box and rack used in thepackaging of disposable pipet tips. The plastic box with a 96-hole rack(8×12 holes) for sterile disposable auto-pipet tips, available from VWRInternational as catalog number 82003-196, STERILE AEROSOL PIPET TIPS 96TIP RACKS, was opened and emptied of the disposable pipet tips. The96-hole rack was removed from the sleeve holder and set aside. Sampledisks of Example 11 and Example 12 (spiked with yeast) were placed onthe sleeve holder. The sleeve holder also contained holes which matchedthe holes in the rack for holding pipet tips. The 96-hole rack was thenreplaced over the samples, thus securing them between the sleeve holderand the rack. This arrangement still exposing most of both sides of thesample disks to the holes in the rack and sleeve holder. The open sideof the sleeve holder was closed by taping the cover of onto the sides ofthe sleeve holder. Monitoring device #2 was created by positioning thesample disks of Example 11 and Example 12 between two stainless steelplates, held in place with typical office supply type paper binderclips. The stainless steel plates each contained an array of 6×6machined holes of about 4 mm in diameter. The disks of Example 11 andExample 12 were positioned such that they were held in place by the twostainless steel plates of Monitoring Device #2 but the majority of thedisks were still exposed on both sides. Both Monitoring device #1 andMonitoring device #2 allowed the free flow of water and detergent overthe sample disks, during the washer disinfection cycle.

Washer Disinfector Cycle

The yeast spiked disks of Example 11 and Example 12, mounted onMonitoring device #1 and Monitoring device #2 were placed the GETINGE46-4 model-washer disinfector (Getinge USA, Inc., Rochester, N.Y.) andsubjected to the INSTRM-LONG-D-3 cycle, a cycle which is commonly usedin hospitals. The optional step of adding “lubricant/instrument milk”during mid cycle was omitted. The washer cycle had both enzymaticdetergent and alkaline detergent as indicated in Table 17, which showsthe steps in the INSTRM-LONG-D-3 wash cycle.

TABLE 17 INSTRM-LONG-D-3 cycle Step Program phase Water Injections Time¹1 Pre-rinse Cold water n/a 3 minutes 2 Wash - 1 Hot water 60 mL Enzyme 7minutes detergent 3 Wash - 2 Hot water 60 mL Alkaline 9 minutesdetergent 4 Post-rinse 1 Hot water n/a 3 minutes 5 Post rinse 2 Hotwater n/a 3 minutes 6 Final rinse Hot water n/a 10 minutes  7 Drying Nowater n/a 12 minutes  ¹All times are approximate and include theequilibration period needed for the washer to reach the prescribedtemperature.

Measurement of ATP from Yeast Spiked CM-111 Carrier Material after WashDisinfection Cycle.

After processing the yeast-spiked disks in the INSTRM-LONG-D-3 washingcycle, the disks of Example 11 and Example 12 were removed fromMonitoring device #1 and Monitoring device #2 and were temporarilytransferred to sterile petri dishes. The disks were then transferredinto empty sterile 1.5 mL polypropylene micro-centrifuge tubes (VWR,Catalog #89000-028). Each of the sample disks were prepared for ATPanalysis by adding 100 microliters of an extractant solution and 500microliters of an enzyme solution from a sample preparation kit(obtained from 3M Company; St. Paul, Minn., under the trade designation3M CLEAN-TRACE SURFACE ATP SYSTEM) to the tube containing the disks. Thecontents were mixed for 5 seconds at about 3200 rpm on a vortex mixer(obtained from VWR, West Chester Pa., under the trade designation VWRFIXED SPEED VORTEX MIXER). The ATP signal of the sample was measured inrelative light units (RLU) for one minute at 10 second intervals using abench-top luminometer (obtained from Turner Biosystems, Sunnyvale,Calif., under the trade designation 20/20N SINGLE TUBE LUMINOMETER,equipped with 20/20n SIS software). Positive control samples were alsocreated by preparing unwashed sample disks of Example 11 and Example 12(spiked with yeast) for ATP analysis in the same fashion as describedabove. The luminescence values in RLU were converted to Log base 10(log₁₀) and are reported in Table 18, below. The Log₁₀ Reduction Value(LRV) was also calculated by subtracting the Log₁₀ RLU value of thewashed disks from the Log₁₀ RLU value of the respective positive control(unwashed disks). All Example results in Table 18 were the average ofduplicate disks. Each average value had a relative variance of less than10%, with the one exception of Example 11 on Monitoring device #1, whichhad a relative variance of 20%. Table 18 demonstrates that a livingmicroorganism such as yeast can be adhered to a carrier and subjected toa wash disinfection cycle to assess the effectiveness of the washdisinfection cycle by measuring the residual amount of ATP.

TABLE 18 Amount of ATP detected from Yeast Spiked Examples 11 and 12.All results are reported in Log₁₀ Relative Light Units (RLU). ATP SignalSample (Log₁₀ RLU) LRV Example 11 Unwashed (positive control) 6.2 —Example 11 on Monitoring device #1 4.6 1.7 Example 11 on Monitoringdevice #2 4.1 2.1 Example 12 Unwashed (positive control) 6.7 — Example12 on Monitoring device #1 3.6 3.1 Example 12 on Monitoring device #24.2 2.6

The complete disclosure of all patents, patent applications, andpublications, and electronically available material cited herein areincorporated by reference. In the event that any inconsistency existsbetween the disclosure of the present application and the disclosure(s)of any document incorporated herein by reference, the disclosure of thepresent application shall govern. The foregoing detailed description andexamples have been given for clarity of understanding only. Nounnecessary limitations are to be understood therefrom. The invention isnot limited to the exact details shown and described, for variationsobvious to one skilled in the art will be included within the inventiondefined by the claims.

All headings are for the convenience of the reader and should not beused to limit the meaning of the text that follows the heading, unlessso specified.

Various modifications may be made without departing from the spirit andscope of the invention. These and other embodiments are within the scopeof the following claims.

1. A monitoring device, comprising: a test composition comprising apredetermined quantity of tracer analyte; a test element comprising atest portion to which the test composition is releasably adhered; adetection reagent; and a container comprising a first end with anopening and a second end opposite the first end; wherein the containeris configured to receive the test portion and configured to beoperationally coupled to an analytical instrument; wherein the traceranalyte and the detection reagent each are capable of participating inone or more chemical reaction that results in the formation of adetectable product. 2.-20. (canceled)